Small molecules against cancer

ABSTRACT

Compounds capable of, or usable in, killing cancer cells, and/or modulating a biological activity of a chemokine, and/or inhibiting a kinase, and/or treating diseases and disorders associated with a biological activity of a chemokine and/or cell migration, and/or treating disease and disorders such as cancer and inflammatory diseases and disorders, are provided herein. The compounds are listed in Tables 2, 4 and 5, and/or are represented by Formulae I, IV, V and VI, as defined in the specification. Methods utilizing these compounds are also provided.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/868,558 filed on May 7, 2020, which is a continuation of U.S. patentapplication Ser. No. 16/063,278 filed on Jun. 17, 2018, now U.S. Pat.No. 10,646,465, which is a National Phase of PCT Patent Application No.PCT/IL2016/051346 having International Filing Date of Dec. 15, 2016,which claims the benefit of priority under 35 USC § 119(e) of U.S.Provisional Patent Application Nos. 62/268,568, 62/268,575 and62/268,579, all filed on Dec. 17, 2015.

The contents of the above applications are all incorporated by referenceas if fully set forth herein in their entirety.

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to therapyand more particularly, but not exclusively, to compounds, compositionsand methods useful in inhibiting a kinase, in modulating a biologicalactivity of a chemokine, in inhibiting cancel cells, in inhibitingchemokine-dependent cell migration and/or in treating diseases anddisorders associated with kinase activity, biological activities ofchemokines and/or cell migration, such as cancer and inflammatorydiseases and disorders.

Chemokines are among the many biological factors that are involved inthe inflammatory disease process. Chemokines belong to a group of small,about 8-14 kDa, mostly basic, heparin-binding proteins that are relatedboth in their primary structure and the presence of 4 conserved cysteineresidues.

The chemokines are chemotactic cytokines that have been shown to beselective chemoattractants for leukocyte sub-populations in vitro, andto elicit the accumulation of inflammatory cells in vivo. In addition tochemotaxis, chemokines mediate leukocyte de-granulation [Baggiolini andDahinden, Immunol Today 1994, 15:127-133], up-regulation of adhesionreceptors [Vaddi and Newton, J Immunol 1994, 153:4721-4732], andsuppression of human immunodeficiency virus replication [Cocchi et al.,Science 1995, 270:1811-1815].

Chemokines play an essential role in the recruitment and activation ofcells from the immune system. They also have a wide range of effects inmany different cell types beyond the immune system, including forexample, in various cells of the central nervous system [Ma et al., PNAS1998, 95:9448-9453], and in endothelial cells, where they result ineither angiogenic or angiostatic effects [Strieter et al., J Biol Chem1995, 270:27348-27357]. Particular chemokines may have multiple effectson tumors, including angiogenesis, promotion of growth and metastasis,and suppression of the immune response to cancer, while other chemokinesinhibit tumor-mediated angiogenesis and promote anti-tumor immuneresponses.

Chemokine receptors have received increasing attention due to theircritical role in the progression of inflammation and associatedconditions such as asthma, atherosclerosis, graft rejection, AIDS andautoimmune conditions (e.g., multiple sclerosis, arthritis, myastheniagravis, lupus).

SDF-1 (stromal cell-derived factor 1), also known as CXCL12 (C-X-C motifchemokine 12), is a chemokine which is strongly chemotactic forlymphocytes. SDF-1 plays an important role in angiogenesis, includingangiogenesis associated with tumor progression, by recruitingendothelial progenitor cells from the bone marrow, an effect mediated bythe CXCR4, the receptor for SDF-1 [Zheng et al., Cardiovasc Pharmacol2007, 50:274-280; Kryczek et al., Am J Physiol Cell Physiol 2007,292:C987-C995]. In addition, cancer cells that express CXCR4 areattracted to metastasis target tissues that release SDF-1.

Plerixafor, an antagonist of CXCR4, is used in combination with G-CSF(granulocyte colony-stimulating factor) to mobilize hematopoietic stemcells in cancer patients, particularly lymphoma and multiple myelomapatients. The stem cells are subsequently transplanted back to thepatient after chemotherapy or radiotherapy.

In animal studies, plerixafor has also been reported to reducemetastasis Smith et al., Cancer Res 2004, 64:8604-8612], to reducerecurrence of glioblastoma associated with vasculogenesis [Kioi et al.,J Clin Investigation 2010, 120:694-705], and to counteractopioid-induced hyperalgesia [Wilson et al., Brain Behav Immun 2011,25:565-573].

Kinases are a family of enzymes that mediate the transfer of a phosphatemoiety from a high energy molecule (such as ATP) to a substrate. Kinasesare involved in many cell-signaling pathways. Protein kinases act onproteins, phosphorylating serine, threonine, tyrosine, or histidineresidues in the protein, and thereby affecting the protein's activity.

Mitogen activated protein kinases (MAPK) constitute a family ofproline-directed serine/threonine kinases that activate their substratesby dual phosphorylation. The p38 MAPKs (p38α, p38β, p38γ and p38δ), forexample, are responsible for phosphorylating and activatingtranscription factors (such as ATF-2, MAX, CHOP and C/ERPb) as well asother kinases (such as MAPKAP-K2/3 or MK2/3), and are themselvesactivated by physical and chemical stress (e.g. UV, osmotic stress),pro-inflammatory cytokines and bacterial lipopolysaccharide (LPS) [Steinet al., Ann Rep Med Chem 1996, 31:289-298; Herlaar & Brown, MolecularMedicine Today 1999, 5:439-447]. The products of p38 phosphorylationhave been shown to mediate the production of pro-inflammatory cytokines.

The implication of kinases pathways on various diseases and disorders,and an anti-inflammatory activity of kinase inhibitors have beendescribed in the art. For example, anti-inflammatory activities havebeen reported for p38 kinase inhibitors [Badger et al., J Pharm ExpThera 1996, 279:1453-1461; Griswold et al., Pharmacol Comm 1996,7:323-229]. In particular, p38 kinase inhibitors have been described aspotential agents for treating rheumatoid arthritis, and to exhibitbeneficial effects in models of airway diseases such as COPD and asthma[Haddad et al, Br J Pharmacol 2001, 132:1715-1724; Underwood et al., AmJ Physiol Lung Cell Mol 2000, 279:895-902; Duan et al., Am J Respir CritCare Med 2005, 171:571-578; Escott et al., Br J Pharmacol 2000,131:173-176; Underwood et al., J Pharmacol Exp Ther 2000, 293:281-288].The implication of the p38MAPK pathway in various diseases has beenreviewed by Chopra et al. [Expert Opinion on Investigational Drugs 2008,17:1411-1425].

The compound8-(2,4-dihydroxy-6-(2-oxoheptyl)-phenoxy)-6-hydroxy-3-pentyl-1H-isochromen-1-onewas isolated from oakmoss, and reported to exhibit potent antibacterialactivity against Legionella, but not against other bacteria [Nomura etal., Biol Pharm Bull 2012, 35:1560-1567].

Arjunolic acid [Ramesh et al., Nat Prod Res 2012, 26:1549-1552],auraptene [Epifano et al., Phytother Res 2013, 27:784-786; Genovese &Epifano, Curr Drug Targets 2011, 1:381-386] and falcarindiol [Huang etal., Molecules 2014, 19:6142-6162; Wang et al., Phytomedicine 2013,20:999-1006] have been reported to inhibit cancer cells. Auraptene hasalso been reported to suppress MCP-1 expression in adipocytes[Kuroyanagi et al., Biochem Biophys Res Commun 2008, 366:219-215].

SUMMARY OF THE INVENTION

According to an aspect of some embodiments of the invention, there isprovided a compound for use in treating cancer, inflammation or anon-cancerous hyperproliferative disorder in a subject in need thereof,the compound being BKT-300, having the formula:

According to an aspect of some embodiments of the invention, there isprovided a compound BKT-300 for use in treating cancer.

According to an aspect of some embodiments of the invention, there isprovided a compound BKT-300 for use in inhibiting SDF-1 and/or CXCR4.

According to an aspect of some embodiments of the invention, there isprovided a compound BKT-300 for use in inducing cell death.

According to an aspect of some embodiments of the invention, there isprovided a compound for use in treating cancer, inflammation or anon-cancerous hyperproliferative disorder in a subject in need thereof,the compound being selected from the group of compounds presented inTable 2 and/or the compound having a general formula selected from thegroup consisting of:

wherein treating a cancer does not comprise illuminating the compound insitu at a wavelength absorbed by the compound of formula IV and/orformula V,

wherein:

W is a hydrocarbon moiety having from 4 to 20 carbon atoms, optionallycomprising one or more hydroxy substituents and/or oxygen atoms betweentwo carbon atoms, wherein a ratio of oxygen atoms to carbon atoms in Wis no more than 1:4;

L₁ is absent or is selected from the group consisting of —O—, —C(═O)—,—C(═O)O—, and —CR₁(OH)—, wherein when L₁ is absent, W is attacheddirectly to X;

X is selected from the group consisting of —C≡C—C≡C—; —CR₃═CR₄—CR₅═CR₆—,a substituted or unsubstituted bicyclic hydrocarbon moiety, andsubstituted or unsubstituted phenylene;

L₂ is absent or is selected from the group consisting of —O—, —C(═O)—,—C(═O)O—, and —C(R₂)(OH)—;

Y is absent or is selected from the group consisting of an aliphatichydrocarbon moiety from 1 to 8 atoms in length, being substituted orunsubstituted, or alternatively, Y attaches to X and/or to Z to form oneor two five- or six-membered rings;

Z is absent or is selected from the group consisting of —C(═O)OH andaryl substituted by at least one hydroxy group; and

R₁-R₆ are each independently selected from the group consisting ofhydrogen, alkyl, alkenyl, alkynyl and cycloalkyl,

wherein when L₂ is absent, Y is absent and Z is —C(═O)OH attacheddirectly to X, and

when Y is absent, L₂ is attached directly to Z, or L₂ is absent and Z is—C(═O)OH attached directly to X;

wherein:

R₄₀-R₅₁ and R₆₁-R₆₈ are each selected from the group consisting ofhydrogen, hydroxy, alkyl, C-carboxy and a saccharide moiety;

R₅₂-R₅₅ are hydrogen, or alternatively, R₅₂ and R₅₃, and/or R₅₄ and R₅₅,are together a covalent bond which forms a six-membered carbon ring; andthe dashed line denotes a saturated or unsaturated bond;

and wherein:

wherein:

A′ is selected from the group consisting of —C(═O)—O—; —NR₇₁—C(═O)—;═C(R₇₂)—N═; —C(R₇₂)═N— and —C(R₇₃)═C(R₇₄)—;

B′ is selected from the group consisting of —NR₇₅—C(═O)—; —C(R₇₆)—N═;—C(R₇₆)═N—; —C(R₇₇)—C(═O)—; and —C(R₇₇)═C(R₇₈)—;

Y′₁ and Y′₂ are each independently selected from C-Q′, C—R₇₉ and N,provided that at least one of Y′₁ and Y′₂ is C-Q′;

Y′₃ is selected from N and C—R₈₀;

Y′₄ is selected from N and C—R₈₁;

R₇₁ and R₇₅ are each independently hydrogen or alkyl;

R₇₂-R₇₄ are each independently selected from hydrogen, alkyl, alkyne,hydroxy, amine, alkoxy, aryloxy, thioalkoxy, thioaryloxy, carbonyl andcarboxylate;

R₇₆-R₇₈ are each independently selected from hydrogen, alkyl, hydroxy,amine, alkoxy, aryloxy, thioalkoxy, thioaryloxy, carbonyl, carboxylate,cinnamic acid, acyl, S(OH)₃ and S—O—O—OH;

R₇₉ is hydrogen or cyano;

R₈₀ and R₈₁ are each independently selected from hydrogen, hydroxy,halo, alkoxy, thioalkoxy, thiol, aryloxy, thioalkoxy, thioaryloxy,carbonyl, amine, and SO₃H; and

Q′ is

wherein:

Z′ is selected from 0, NH, C(═O), S, CH₂ and S(═O);

X′₁ is C(R₈₂) or N;

X′₂ is C(R₈₃) or N;

X′₃ is C(R₈₄) or N;

X′₄ is C(R₈₅) or N; and

X_(′5) is C(R₈₆) or N,

provided that at least two of X₇₁-X₇₅ are not N,

R₈₂ is selected from hydrogen, alkyl and halo; and

R₈₃-R₈₆ are each independently selected from hydrogen, halo, alkyl,amine, alkoxy, aryloxy and hydroxy;

or, alternatively, two of R₈₂-R₈₆ are joined together to form an aryl orheteroaryl.

According to an aspect of some embodiments of the invention, there isprovided a compound for use in modulating a biological activity of achemokine in a subject in need thereof, the compound being selected fromthe group of compounds presented in Table 2 and/or the compound having ageneral formula selected from the group consisting of Formulae I, IV, Vand VI, as defined herein in any of the respective embodiments.

According to an aspect of some embodiments of the invention, there isprovided a compound for use in inducing cell death in a subject in needthereof, the compound being selected from the group of compoundspresented in Table 2 and/or the compound having a general formulaselected from the group consisting of Formulae I, IV, V and VI, asdefined herein in any of the respective embodiments.

According to an aspect of some embodiments of the invention, there isprovided a compound for use in inhibiting a kinase and/or in treating adisease or disorder associated with an activity of a kinase, thecompound being represented by formula VI, as defined herein in any ofthe respective embodiments.

According to an aspect of some embodiments of the invention, there isprovided a compound being represented by Formula VIA:

wherein:

A′ is selected from the group consisting of —C(═O)—O— and —NR₁—C(═O)—;

B′ is selected from the group consisting of —NR₇₅—C(═O)—;—C(R₇₇)—C(═O)—; and —C(R₇₇)═C(R₇₈)—;

Y′₁ and Y′₂ are each independently selected from C-Q′, CH and N,provided that at least one of Y′₁ and Y′₂ is C-Q′;

R₇₁ and R₇₅ are each independently hydrogen or alkyl; and

R₇₇ and R₇₈ are each independently hydrogen and alkyl,

at least one of R₇₁, R₇₇ and R₇₈ is an alkyl being at least 4 atoms inlength;

R₈₀ and R₈₁ are each independently selected from hydrogen, alkoxy,aryloxy and hydroxyl, at least one R₈₀ and R₈₁ being hydroxyl or alkoxy;and

Q′ is

wherein:

Z′ is O;

X₁ is C(R₈₂);

X′₂ is C(R₈₃) or N;

X′₃ is C(R₈₄);

X′₄ is C(R₈₅) or N; and

X′₅ is C(R₈₆) or N,

R₈₂ is selected from hydrogen and alkyl; and

R₈₃-R₈₆ are each independently selected from hydrogen, alkoxy, aryloxyand hydroxyl, at least one of R₈₃-R₈₆ being independently hydroxyl,alkoxy or aryloxy.

According to some embodiments of any one of the embodiments of theinvention relating to cancer, the cancer is selected from the groupconsisting of a leukemia, a lymphoma and a lung cancer.

According to some embodiments of any one of the embodiments of theinvention relating to cancer, the cancer is selected from the groupconsisting of acute myeloid leukemia, acute lymphoblastic leukemia,Burkitt lymphoma, multiple myeloma, large cell lung cancer and smallcell lung cancer.

According to some embodiments of any one of the embodiments of theinvention relating to cancer, the cancer is characterized by expressionof CXCR4.

According to some embodiments of any one of the respective embodimentsof the invention, treating a cancer further comprises administering anadditional anti-cancer agent.

According to some embodiments of the invention, the additionalanti-cancer agent is selected from the group consisting ofcombretastatin A-4 phosphate and ombrabulin.

According to some embodiments of any one of the respective embodimentsof the invention, treating a cancer increases a level of hematopieticstem cells in peripheral blood of the subject, and further comprisesobtaining hematopietic stem cells from peripheral blood of the subject,administering a cytotoxic therapy to the subject, and transplanting thestem cells back into the subject subsequent to the cytotoxic therapy.

According to some embodiments of any one of the embodiments of theinvention relating to inhibiting SDF-1 and/or CXCR4, the compound is foruse in treating a disease or disorder associated with an activity ofSDF-1 and/or CXCR4.

According to some embodiments of the invention, the disease or disorderassociated with an activity of SDF-1 and/or CXCR4 is selected from thegroup consisting of harmful angiogenesis, tumor metastasis, WHIMsyndrome and opioid-induced hyperalgesia.

According to some embodiments of any one of the respective embodimentsof the invention, inhibiting SDF-1 and/or CXCR4 is for effectingimmunostimulation.

According to some embodiments of any one of the embodiments of theinvention relating to modulating a chemokine activity, the activity ischemokine induced cell migration.

According to some embodiments of any one of the respective embodimentsof the invention, the chemokine is selected from the group consisting ofMIP3a, MCP-1 and SDF-1.

According to some embodiments of any one of the embodiments of theinvention relating to modulating a chemokine activity, the compound isfor use in inhibiting a biological activity of SDF-1 and/or CXCR4.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, the compound is for use intreating a cancer, wherein: X is selected from the group consisting of—CR₃═CR₄—CR₅═CR₆—, a substituted or unsubstituted bicyclic hydrocarbonmoiety and substituted or unsubstituted phenylene; and

L₂ is absent or is selected from the group consisting of —C(═O)— and—C(R₂)(OH)—.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, R₁ and R₂ are eachhydrogen.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, the phenylene issubstituted by at least one substituent selected from the groupconsisting hydroxy and a saccharide moiety.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, R₃-R₆ are each hydrogen.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, when Y attaches to X toform one or two five- or six-membered rings, Y is selected from thegroup consisting of —CH═CR₇-A- and —CH₂—CHR₈—B—,

wherein:

A and B are each independently absent or an oxygen atom attached to X orto L₂, wherein when A or B is attached to L₂, L₂ is —C(═O)—; and

R₇ and R₈ are each independently selected from the group consisting ofhydrogen, alkyl and Z.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, at least one ring atom ofeach of the five- or six-membered rings is an oxygen atom.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, Z is selected from thegroup consisting of hydroxyphenyl, dihydroxyphenyl and —C(═O)OH.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, W comprises a substitutedor unsubstituted alkyl or alkenyl group, the substituted orunsubstituted alkyl or alkenyl comprising at least 4 carbon atoms.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, the one or more oxygenatoms in W are each within a hydroxy or furanyl group.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula I, L₁ is absent or isselected from the group consisting of —O—, —C(═O)—, and —CR₁(OH)—.

According to some embodiments of the invention, the compound is acompound having the general formula II:

wherein:

each of the dashed lines independently denotes a saturated orunsaturated bond;

J is selected from the group consisting of —O— and —CR₁₄═;

K is selected from the group consisting of —C(═O)—, —CR₁₅═ and—CR₁₆R₁₇—;

M is selected from the group consisting of —O—, —CR₁₈═ and —CR₁₉R₂₀—;

Q is selected from the group consisting of —C(═O)— and —CR₂₁═;

R₁₀, R₁₂ and R₁₃ are each independently selected from the groupconsisting of hydrogen, hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl,alkoxy and aryloxy;

R₁₁ is selected from the group consisting of hydrogen, alkyl, alkenyl,alkynyl, cycloalkyl and heteroalicyclic, or alternatively, R₁ and R₁₂together form a five- or six-membered heteroaryl or heteroalicyclicring;

R₁₄-R₂₁ are each independently selected from the group consisting ofhydrogen, alkyl, alkenyl, alkynyl, cycloalkyl and aryl; and and at leastone of K and Q is —C(═O)—, and at least one of J and M is —O—.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula II, no more than one of K andQ is —C(═O)—.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula II, no more than one of J andM is —O—.

According to some embodiments of the invention, the compound is acompound having the general formula III:

wherein:

D is selected from the group consisting of —CH₂— and —C(═O)—; the dashedlines each denote a saturated or unsaturated bond;

p is an integer in a range of from 0 to 6;

R₃₀ and R₃₁ are each hydrogen or R₃₀ and R₃₁ to together form anaromatic ring substituted by at least one hydroxy group, wherein whenR₃₀ and R₃₁ are each hydrogen, D is —C(═O)— and each of the dashed linesdenotes an unsaturated bond; and

R₃₂ is selected from the group consisting of an alkyl, alkenyl and acyl,each being from 2 to 12 carbon atoms in length.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula IV and/or V, R₄₀-R₅₁ andR₆₁-R₆₈ are each selected from the group consisting of hydrogen,hydroxy, alkyl and alkylcarboxy.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula IV:

the dashed line in Formula IV denotes an unsaturated bond, R₅₂ and R₅₃are together a covalent bond which forms a six-membered aromatic carbonring, and R₅₄ and R₅₅ are together a covalent bond which forms asix-membered aromatic carbon ring; or

the dashed line in Formula IV denotes a saturated bond, and R₅₂-R₅₅ areeach hydrogen.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VI:

A′ is selected from the group consisting of —C(═O)—O—; and —NR₇₁—C(═O)—;

B′ is selected from the group consisting of —NR₇₅—C(═O)—;—C(R₇₇)—C(═O)—; and C(R₇₇)═C(R₇₈);

Y′₁ and Y′₂ are each independently selected from C-Q′, C—R₇₉ and N,provided that at least one of Y′₁ and Y′₂ is C-Q′;

Y′₃ is C—R₈₀;

Y′₄ is C—R₈₁;

R₇₁ and R₇₅ are each independently hydrogen or alkyl;

R₇₇ and R₇₈ are each independently selected from hydrogen and alkyl,

wherein at least one of R₇₁, R₇₇ and R₇₈ is an alkyl being at least 4atoms in length;

R₇₉ is hydrogen;

R₈₀ and R₈₁ are each independently selected from hydrogen, alkoxy andhydroxyl, at least one of R₈₀ and R₈₁ being hydroxyl or alkoxy; and

Q′ is

wherein:

Z′ is O;

X′₁ is C(R₈₂);

X′₂ is C(R₈₃) or N;

X′₃ is C(R₈₄);

X′₄ is C(R₈₅) or N; and

X′₅ is C(R₈₆) or N,

provided that at least two of X₇₁-X₇₅ are not N,

R₈₂ is selected from hydrogen and alkyl; and

R₈₃-R₈₆ are each independently selected from hydrogen, alkoxy andhydroxyl, at least one of R₈₃-R₈₆ being hydroxyl or alkoxy.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VI:

A′ is C(═O)—O—; and

B′ is C(R₇₇)═C(R₇₈),

and wherein R₇₇ is the alkyl being at last 4 carbon atoms in length.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VI:

A′ is —NR₇₁—C(═O)—; and

B′ is —NR₇₅—C(═O)—,

and wherein R₇₁ is the alkyl being 4 carbon atoms in length.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VI, each of X₇₁-X₇₅ is otherthan N.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VI, at least one of X₇₂, X₇₄or X₇₅ is N.

According to some embodiments of the invention, the compound is selectedfrom the compounds presented in Table 4 and/or in Table 5.

According to some embodiments of the invention, the compound is selectedfrom the compounds presented in Table 2.

According to some embodiments of the invention, the compound selectedfrom the group consisting of BKT206, BKT211, BKT215 and BKT300 in Table2, the compound being for use in treating a cancer.

According to some embodiments of any one of the embodiments of theinvention relating to inhibiting a kinase, the compound is BKT-300.

According to some embodiments of any one of the embodiments of theinvention relating to inhibiting a kinase, the kinase is selected fromthe group consisting of DYRK3, EPHA8, GRK4, GRK5, MAP4K1, MAP4K2,MAP4K4, MELK, PAK7, SGK2, SRC N1, ACVRL1, BMPRA, CDCl₇/DBF4, CDK1/cyclinA2, CDK11, CDK8/cyclin C, CLK4, DAPK2, DURK2, ICK, MAPK10, MLCK, MYLK,NUAK2, STK17A, STK17B, STK38, STK38L, TGFBR2, TTK, DAPK1, PIK3CA andPIK3CD.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VIA:

A′ is C(═O)—O—; and

B′ is C(R₇₇)═C(R₇₈),

and wherein R₇₇ is the alkyl being at last 4 carbon atoms in length.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VIA:

A′ is —NR₇₁—C(═O)—; and

B′ is —NR₇₅—C(═O)—,

and wherein R₇₁ is the alkyl being 4 carbon atoms in length.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VIA, at least one of X′₂, X′₄or X′₅ is N.

According to some embodiments of any one of the embodiments of theinvention relating to compounds of Formula VIA, the compound is selectedfrom the compounds presented in Table 5.

Unless otherwise defined, all technical and/or scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which the invention pertains. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of embodiments of the invention, exemplarymethods and/or materials are described below. In case of conflict, thepatent specification, including definitions, will control. In addition,the materials, methods, and examples are illustrative only and are notintended to be necessarily limiting.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

Some embodiments of the invention are herein described, by way ofexample only, with reference to the accompanying drawings. With specificreference now to the drawings in detail, it is stressed that theparticulars shown are by way of example and for purposes of illustrativediscussion of embodiments of the invention. In this regard, thedescription taken with the drawings makes apparent to those skilled inthe art how embodiments of the invention may be practiced.

In the drawings:

FIG. 1 is a graph showing effects of 3,500 compounds (black dots) onbinding of BKT130 to MIP3a, as determined by high-throughput screening,as well as control samples (blue dots) in which MIP3a is absent; x-axisshows sample numbers, whereas y-axis values represent binding, whereinzero represents no effect on binding, and −100 represents absence ofbinding (full inhibition, or absence of MIP3a).

FIGS. 2A-2C are graphs showing the activity (inhibition of binding ofBKT130 to MIP3a) of 3 exemplary compounds (one in each of FIGS. 2A-2C)as a function of concentration.

FIG. 3 is a bar graph showing the effect of Compound BKT300 (at 78%purity) (at concentrations of 10 and 50 μg/ml) on migration of CD4+cells towards MIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 4 is a bar graph showing the effect of Compound BKT201 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 5 is a bar graph showing the effect of Compound BKT202 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 6 is a bar graph showing the effect of Compound BKT205 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 7 is a bar graph showing the effect of Compound BKT209 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 8 is a bar graph showing the effect of Compound BKT210 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 9 is a bar graph showing the effect of Compound BKT213 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.05 vs. zero concentration).

FIG. 10 is a bar graph showing the effect of Compound BKT203 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.01 vs. zero concentration).

FIG. 11 is a bar graph showing the effect of Compound BKT206 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.01 vs. zero concentration).

FIG. 12 is a bar graph showing the effect of Compound BKT207 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.01 vs. zero concentration).

FIG. 13 is a bar graph showing the effect of Compound BKT212 (atconcentrations of 10 and 50 μg/ml) on migration of CD4+ cells towardsMIP3a (* indicates p<0.01 vs. zero concentration).

FIGS. 14A and 14B present bar graphs showing the effect of 10 and 50μg/ml BKT300 (at 78% purity) on migration of Jurkat cells towards SDF-1(FIG. 14A), and the effect of 1 and 10 μg/ml of Compound BKT300 (at 98%purity) on migration of Jurkat cells towards SDF-1 (* indicates p<0.05vs. zero concentration).

FIG. 15 is a bar graph showing the effect of 1 and 10 μg/ml of CompoundBKT400 on migration of Jurkat cells towards SDF-1 (* indicates p<0.05vs. zero concentration).

FIG. 16 is a bar graph showing the effect of Compound BKT201 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.01 vs. zero concentration).

FIG. 17 is a bar graph showing the effect of Compound BKT203 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 18 is a bar graph showing the effect of Compound BKT204 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 19 is a bar graph showing the effect of Compound BKT205 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.01 vs. zero concentration).

FIG. 20 is a bar graph showing the effect of Compound BKT206 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 21 is a bar graph showing the effect of Compound BKT207 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 22 is a bar graph showing the effect of Compound BKT208 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 23 is a bar graph showing the effect of Compound BKT209 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.01 vs. zero concentration).

FIG. 24 is a bar graph showing the effect of Compound BKT210 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 25 is a bar graph showing the effect of Compound BKT211 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 26 is a bar graph showing the effect of Compound BKT212 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.05 vs. zero concentration).

FIG. 27 is a bar graph showing the effect of Compound BKT213 (atconcentrations of 10 and 50 μg/ml) on migration of Jurkat cells towardsSDF-1 (* indicates p<0.01 vs. zero concentration).

FIG. 28 is a bar graph showing the effect of 10 and 50 μg/ml of CompoundBKT300 (at 78% purity) on migration of THP-1 cells towards MCP-1 (*indicates p<0.05 vs. zero concentration).

FIG. 29 is a bar graph showing the effect of Compound BKT201 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.05 vs. zero concentration).

FIG. 30 is a bar graph showing the effect of Compound BKT204 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.05 vs. zero concentration).

FIG. 31 is a bar graph showing the effect of Compound BKT205 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.05 vs. zero concentration).

FIG. 32 is a bar graph showing the effect of Compound BKT206 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.05 vs. zero concentration).

FIG. 33 is a bar graph showing the effect of Compound BKT209 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.05 vs. zero concentration).

FIG. 34 is a bar graph showing the effect of Compound BKT211 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.01 vs. zero concentration).

FIG. 35 is a bar graph showing the effect of Compound BKT216 (atconcentrations of 10 and 50 μg/ml) on migration of THP-1 cells towardsMCP-1 (* indicates p<0.05 vs. zero concentration).

FIGS. 36A and 36B are bar graphs showing the effect of 0, 0.1, 1 and 10μg/ml of Compound BKT206 on the viability of MV4-11 cells, as expressedby percentage of dead cells (FIG. 36A) and the number of viable cells(FIG. 36B), as determined by propidium iodide staining (* indicatesp<0.05 vs. zero concentration).

FIGS. 37A and 37B are bar graphs showing the effect of 0, 0.1, 1 and 10μg/ml of Compound BKT211 on the viability of MV4-11 cells, as expressedby percentage of dead cells (FIG. 37A) and the number of viable cells(FIG. 37B), as determined by propidium iodide staining (* indicatesp<0.05 vs. zero concentration).

FIGS. 38A and 38B are bar graphs showing the effect of 0, 5, 10 and 20μM (equivalent to 0, 1.5, 3 and 6 μg/ml) of BKT215 on the viability ofMV4-11 cells, as expressed by percentage of dead cells (FIG. 38A) andthe number of viable cells (FIG. 38B), as determined by propidium iodidestaining (* indicates p<0.05 vs. zero concentration).

FIGS. 39A and 39B are bar graphs showing the effect of 0, 2, 4, 6, 8 and10 μg/ml of BKT300 (at 78% purity) on the viability of MV4-11 cells, asexpressed by percentage of dead cells (FIG. 39A) and the number ofviable cells (FIG. 39B), as determined by propidium iodide staining (*indicates p<0.05 vs. zero concentration).

FIGS. 40A-40D are bar graphs showing the effect of 0, 2.125, 4.25 and8.5 μg/ml (equivalent to 0, 5, 10 and 20 μM) of BKT300 at 78% purity(FIGS. 40A and 40B) and at 98% purity (FIGS. 40C and 40D) on theviability of MV4-11 cells, as expressed by percentage of dead cells(FIGS. 40A and 40C) and the number of viable cells (FIGS. 40B and 40D),as determined by propidium iodide staining (* indicates p<0.05 vs. zeroconcentration).

FIGS. 41A and 41B are bar graphs showing the effect of 0, 2, 4, 6, 8 and10 μg/ml of BKT300 (at 78% purity) on the viability of RPMI cells, asexpressed by percentage of dead cells (FIG. 41A) and the number ofviable cells (FIG. 41B), as determined by propidium iodide staining (*indicates p<0.05 vs. zero concentration).

FIGS. 42A and 42B are bar graphs showing the effect of 0, 5, 10 and 20μM (equivalent to 0, 2.125, 4.25 and 8.5 μg/ml) of BKT300 (at 78%purity) on the viability of Jurkat cells, as expressed by percentage ofdead cells (FIG. 42A) and the number of viable cells (FIG. 42B), asdetermined by propidium iodide staining (* indicates p<0.05 vs. zeroconcentration).

FIGS. 43A and 43B are bar graphs showing the effect of 0, 5, 10 and 20μM (equivalent to 0, 2.125, 4.25 and 8.5 μg/ml) of BKT300 (at 78%purity) on the viability of Raji cells, as expressed by percentage ofdead cells (FIG. 43A) and the number of viable cells (FIG. 43B), asdetermined by propidium iodide staining (* indicates p<0.05 vs. zeroconcentration).

FIGS. 44A and 44B are bar graphs showing the effect of 0, 5, 10 and 20μM (equivalent to 0, 2.125, 4.25 and 8.5 μg/ml) of BKT300 (at 78%purity) on the viability of Bjab cells, as expressed by percentage ofdead cells (FIG. 44A) and the number of viable cells (FIG. 44B), asdetermined by propidium iodide staining (* indicates p<0.05 vs. zeroconcentration).

FIGS. 45A and 45B are bar graphs showing the effect of 0, 0.1, 1 and 10μg/ml of BKT300 (at 78% purity) on the viability of H-460 cells, asexpressed by percentage of dead cells (FIG. 45A) and the number ofviable cells (FIG. 45B), as determined by propidium iodide staining (*indicates p<0.05 vs. zero concentration).

FIGS. 46A and 46B are bar graphs showing the effect of 0, 5, 10 and 20μM (equivalent to 0, 2.125, 4.25 and 8.5 μg/ml) of BKT300 (at 78%purity) on the viability of H345 cells, as expressed by percentage ofdead cells (FIG. 46A) and the number of viable cells (FIG. 46B), asdetermined by propidium iodide staining (* indicates p<0.05 vs. zeroconcentration).

FIGS. 47A-47C present bar graphs showing the effect of intraperitonealadministration of BKT300 (at 98% purity) on the percentage ofCD45-positive cells in the bone marrow of mice injected with 10×10⁶MV4-11 cancer cells 21 days before administration of BKT300 (FIG. 47A),and data of the FACS analysis showing human MV4-11 cancer cells in thebone marrow of untreated (FIG. 47B) and treated with BKT300 (FIG. 47C)mouse 21 days following transplantation of 10×10⁶ MV4-11 cancer cells.

FIG. 48 presents a scheme showing the principles of a FRET assay fordetermining binding of a compound (inhibitor) to an active site ofkinases, wherein resonant energy transfer of energy from a europium(Eu)-labeled antibody which binds to the kinase to an AlexaFluor®-labeled tracer which binds to the active site is prevented by acompound (inhibitor) which binds to the active site.

FIG. 49 presents an illustration of the alignment of MELK and MAPK4Kactive sites; MELK is shown in blue (PDB 4BKY); MAPK4K is shown in green(PDB 40BQ); the small molecule is an inhibitor of MAPK4K (PDB 40BQ).

FIG. 50 is an illustration of BKT300 docked into the ATP binding pocketof MELK.

FIG. 51 is an illustration showing BKT300 (in pink) overlaid on arepresentative small molecule inhibitor of MAPK4K (PDB 40BQ; in green),and a representative small molecule inhibitor of MELK (PDB 4BKY; inblue); the atoms of the known inhibitors that are close to the aliphatictails of BKT300 are marked as balls.

FIGS. 52A-52B present schemes depicting an exemplary synthesis ofBKT300-7 (FIG. 52B) and of a reactant R11 usable in the synthesis (FIG.52A), according to some embodiments of the present invention.

FIGS. 53A-53B present schemes depicting an exemplary synthesis ofBKT300-23 (FIG. 53B) and of a reactant R1 usable in the synthesis (FIG.53A), according to some embodiments of the present invention.

FIG. 54 presents schemes depicting an exemplary synthesis of BKT300-1,according to some embodiments of the present invention.

FIG. 55 presents schemes depicting an exemplary synthesis of BKT300-3,according to some embodiments of the present invention.

FIG. 56 presents schemes depicting an exemplary synthesis of BKT300-11,according to some embodiments of the present invention.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention, in some embodiments thereof, relates to therapyand more particularly, but not exclusively, to compounds, compositionsand methods useful in inhibiting a kinase, in modulating a biologicalactivity of a chemokine, in inhibiting cancel cells, in inhibitingchemokine-dependent cell migration and/or in treating diseases anddisorders associated with kinase activity, biological activities ofchemokines and/or cell migration, such as cancer and inflammatorydiseases and disorders.

Before explaining at least one embodiment of the invention in detail, itis to be understood that the invention is not necessarily limited in itsapplication to the details set forth in the following description orexemplified by the Examples. The invention is capable of otherembodiments or of being practiced or carried out in various ways.

In a search for compounds suitable for modulating chemokine activity,and treating conditions associated with the biological activity ofchemokines, the present inventors have screened a library of about 3,500natural compounds for chemokine-binding activity, and then furtherscreened the chemokine-binding molecules for an ability to modulate theeffect of individual chemokines on cells, as well as for an ability toaffect cancer cells (e.g., by killing cancer cells, inhibiting growth ofcancer cells and/or inhibiting chemokine-dependent migration of cancercells) and/or kill pathogenic cells such as cancer cells.

Using this laborious screening process, the present inventors haveuncovered that compounds characterized by certain structural features,as detailed herein, were shown to exhibit an ability to modulatechemokine activity and/or inhibit cancer cells.

Exemplary such compounds, which were identified through the laboriousscreening process as chemokine-binding small molecules, are presented bytheir chemical structures in Table 2 in the Examples section herein.Many of said compounds surprisingly exhibits relatively selectiveinhibition of SDF-1/CXCR4 activity and/or induction of cancer celldeath. For example, the present inventors have identified the compoundreferred to herein as BKT300 (see the Examples section that follows) asa promising modulator of chemokine activity, a selective inhibitor ofSDF-1/CXCR4 activity and/or as inducing cancer cell death.

Reference is made to FIGS. 2A-13, which show inhibition of the chemokineMIP3a by exemplary chemokine-binding compounds according to embodimentsof the present invention. FIGS. 14A-27 show inhibition by exemplarychemokine-binding compounds of cell migration towards the chemokineSDF-1. FIGS. 28-35 show inhibition by exemplary chemokine-bindingcompounds of cell migration towards the chemokine MCP-1. FIGS. 3, 4, 6,7, 9, 14A, 14B, 16, 19, 23, 27-29, 31 and 33 show that some exemplaryinhibitors of SDF-1 are relatively selective inhibitors of SDF-1.

Further reference is made to FIGS. 36A-46B, which show that CompoundsBKT206, BKT211, BKT215 and BKT300 induce cell death in cancer cells. Inaddition, FIGS. 47A-47C shows that BKT300 reduces cancer cell numbers inan in vivo mouse model.

The small molecule BKT300 was also screened for its activity on aselected list of human kinases and was shown to inhibit activity ofcertain kinases (see, Table 3, Example 5, in the Examples section thatfollows).

Encouraged by the pronounced activity of BKT300, the present inventorshave studied the interactions of BKT300 with the binding site ofkinases, using computational modeling (see, for example, FIGS. 49-51),and based on the data retrieved in these computational study, havedesigned small molecules that are structural analogs of BKT300, whichmaintain the structural features of BKT300 which were considered asattributing to its activity (see, for non-limiting examples, FIGS.52A-56). The present inventors have further identified small moleculeswhich feature a structural topology similar to BKT300, and which can beuseful in inhibiting a kinase activity and/or in treating a disease ordisorder in which inhibition of kinase is beneficial. The smallmolecules described herein are further useful in modulating a biologicalactivity of chemokines and accordingly in treating a disease or disorderthat is associated with a biological activity of a chemokine. The smallmolecules described herein are particularly useful as anti-canceragents, by inducing cancer cells death (e.g., via apoptotic cell death)and/or effecting cancel cells migration (by inhibiting angiogenesisand/or metastasis), as described in further detail hereinbelow. Thesmall molecules described herein are further useful in treatinginflammation (e.g., in treating inflammatory diseases and disorders asdescribed herein). The small molecules described herein are furtheruseful in treating non-cancerous proliferative diseases, as describedherein. Embodiments of the present invention therefore generally relateto newly designed small molecules and to uses thereof.

Embodiments of the present invention further relate to a method ofidentifying a compound (small molecule) usable in any of the usesdescribed herein, which method is also referred to herein as a screeningassay or method.

Compounds:

The compounds described in some embodiments of any of the aspects of thepresent embodiments, and any combination thereof are characterized by arelatively rigid core hydrocarbon moiety, which is preferably cyclicand/or unsaturated, attached directly to one or more small, relativelypolar oxygen-containing groups (for example, —C(═O)—, —CH(OH)— or—C(═O)OH). The core hydrocarbon moiety is further attached to a secondhydrocarbon moiety having at least 4 carbon atoms, which is preferablyrelatively non-polar and at least partially aliphatic. The corehydrocarbon moiety may be attached to the second hydrocarbon moietydirectly or via one of the abovementioned small, oxygen-containinggroups.

The compounds described in some embodiments of any of the aspects of thepresent embodiments, and any combination thereof are, are collectivelyrepresented by Formula I:

W-L₁-X-L₂-Y—Z  Formula I

wherein X in Formula I is a rigid hydrocarbon moiety.

Herein, a “rigid” moiety is a moiety comprising a backbone in which nomore than one bond within the backbone (i.e., a bond between two atomswhich are each part of the backbone) is a free-to-rotate bond (asdefined herein) in which rotation of the free-to-rotate bond affects therelative positions of at least one these moieties, and in which at leastone bond within the backbone is not a free-to-rotate bond. For example,rotation of the single bond in a C≡C—C≡C moiety does not affect therelative position of any of the atoms in the moiety, even though it is afree-to-rotate bond (as defined herein).

A “backbone” of X in Formula I is a chain of carbon atoms in which thecarbon atom at one end of the chain is attached to W-L₁- and the carbonatom at the other end of the chain is attached to -L₂-Y—Z.

The phrase “free-to-rotate bond”, as used herein, describes a bond thatconnects two moieties in a compound, the bond capable of rotating aroundan axis, whereby such a rotation affects the relative positions of thesemoieties, without requiring simultaneous rotation around the axis ofanother bond. A free-to-rotate bond includes a single (sigma) bond whichhas an ability to rotate along its axis, and which is not a bond betweentwo atoms which both form a part of the same ring.

In some embodiments, the rigid moiety comprises one or more unsaturatedbonds (which are not free-to-rotate bonds). In some embodiments, therigid moiety comprises two or more unsaturated bonds which are coplanar.

The rigid core X can be, for example, and unsaturated linear alkylenechain which comprise one or more moeities such as, but not limited to,—C≡C—C≡C—; —CR₃═CR₄—CR₅═CR₆—. Alternatively, the rigid core X can be asubstituted or unsubstituted bicyclic hydrocarbon moiety, or asubstituted or unsubstituted phenylene, as described in more detailhereinafter.

Herein, a moiety (e.g., an X moiety) may be considered “unsubstituted”when attached to a neighboring moiety in a formula described herein(e.g., a W, L₁, L₂ or Z moiety in Formula I), that is, the moietiesdescribed herein are not considered herein to be substituents.

The core moiety represented by X is attached to a relatively polaroxygen-containing group represented by the linking group L₂.Alternatively, X is attached to an oxygen-containing group which is aterminal group (e.g., —C(═O)OH) rather than a linking group, in whichcase, the oxygen-containing group is represented by variable Z inFormula I, and L₂ and Y are absent. X is optionally attached to one ormore additional relatively polar oxygen-containing groups, representedby the optional linking moiety L₁ and/or in one or more substituents ofX.

In embodiments wherein L₂ is not absent, L₂ can be, for example (withoutlimitation), —O—, C(═O)—, —C(═O)O— (wherein either the carbon atom oroxygen atom may be attached to X), and/or —C(R₂)(OH)—. In someembodiments, L₂ is absent or is —C(═O)— or —C(R₂)(OH)—. In someembodiments, L₂ is absent or is —C(═O)—.

In some embodiments, L₂ is absent, Y is absent and Z is a terminal polaroxygen-containing group (e.g., as described herein for L₂) which isattached directly to X. In some of these embodiments, Z is —C(═O)OH.

L₁ in Formula I can be absent or an optional oxygen-containing linkinggroup, as described herein for L₂. In embodiments where L₁ is notabsent, it can be, for example, —O—, —C(═O)—, —C(═O)O— (wherein eitherthe carbon atom or oxygen atom may be attached to X), and/or —CR₁(OH)—.In some embodiments, L₁ is absent or is —O—, —C(═O)—, and/or —CR₁(OH)—.In embodiments wherein L₁ is absent, W is attached directly to X.

The abovementioned substituents R₁-R₆ can be, for example, hydrogen,alkyl, alkenyl, alkynyl and/or cycloalkyl.

In some embodiments of any of the embodiments described herein relatingto Formula I, R₁ is hydrogen.

In some embodiments of any of the embodiments described herein relatingto Formula I, R₂ is hydrogen. In some such embodiments, R₁ and R₂ areeach hydrogen.

W in Formula I is a hydrocarbon moiety having from 4 to 20 carbon atoms,which is attached to X directly or via the L₁ linking group. Thehydrocarbon moiety optionally comprises one or more hydroxy substituentsand/or oxygen atoms between two carbon atoms (e.g., interrupting ahydrocarbon chain), wherein a ratio of oxygen atoms to carbon atoms in Wis no more than 1:4 (oxygen atoms:carbon atoms). In some embodiments, Wcomprises from 5 to 20 carbon atoms. In some embodiments, W comprisesfrom 5 to 15 carbon atoms.

As used herein throughout, the term “hydrocarbon” describes an organicmoiety that includes, as its basic skeleton, a chain of carbon atoms,substituted mainly or entirely by hydrogen atoms. The hydrocarbon can besaturated or non-saturated, be comprised of aliphatic, alicyclic oraromatic moieties, and can optionally (unless indicated otherwise) besubstituted by one or more substituents (other than hydrogen). Thehydrocarbon moiety is optionally interrupted by one or more oxygen atoms(unless indicated otherwise). When substituted, the substituent groupcan be, for example, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy,aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl,cyano, nitro, azide, phosphonyl, phosphinyl, oxo, thiocarbonyl, urea,thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, and amine, as theseterms are defined herein.

The W moiety is attached to X or L₁ via a covalent bond, oralternatively, W comprises a cyclic (e.g., substituted or nonsubstitutedcycloalkyl or aryl moiety) which is fused to a ring of the X moiety(i.e., to a phenylene ring or one or both rings of a bicyclichydrocarbon moiety, according to any of the respective embodimentsdescribed herein), and L₁ is absent. In some such embodiments, Wcomprises a substituted or unsubstituted cycloalkyl group fused to X. Insome such embodiments, X is a saturated or unsaturated aliphaticbicyclic hydrocarbon moiety (according to any of the respectiveembodiments described herein). Compound BKT216 is an example of acompound comprising a cycloalkyl group of the W moiety fused to a ringof an X moiety (which is a saturated aliphatic bicyclic moiety).

In some embodiments of any of the embodiments described herein relatingto Formula I, W is attached to L₁ or X via a covalent bond, that is, Wis not fused to X. In some such embodiments, L₁ is absent and W isattached to X directly via a covalent bond.

Without being bound by any particular theory, it is believed that themoiety denoted as W in Formula I interacts with a substrate (e.g., achemokine) via hydrophobic interactions. Accordingly, in someembodiments, W is a hydrophobic moiety, and in some such embodiments,the number of oxygen moieties moiety is such that do not affect itscapability of participating in hydrophobic interactions.

In some embodiments of any of the embodiments described herein relatingto Formula I, a ratio of oxygen atoms to carbon atoms in the moietydenoted as W is no more than 1:5 (oxygen atoms:carbon atoms). Thus, forexample, when W contains 4 carbon atoms, no oxygen atoms are present inW. In some such embodiments, the ratio is no more than 1:6. In some suchembodiments, the ratio is no more than 1:7. In some such embodiments,the ratio is no more than 1:8. In some such embodiments, the ratio is nomore than 1:9. In some such embodiments, the ratio is no more than 1:10.In some such embodiments, the ratio is no more than 1:12. In some suchembodiments, W is devoid of oxygen atoms.

In some embodiments of any of the embodiments described herein relatingto Formula I, an oxygen atom between two carbon atoms in W is within afuran heteroaryl ring. In some such embodiments, each oxygen atom in Wis within a hydroxy or furanyl (optionally unsubstituted furanyl) group.In some such embodiments, each oxygen atom in W is within a furan ring(e.g., unsubstituted furanyl). It is to be appreciated that furan groupsare relatively hydrophobic in comparison to other oxygen containinggroups.

In some embodiments of any of the embodiments described herein relatingto Formula I, W comprises at least one substituted or unsubstitutedalkyl or alkenyl group. In some such embodiments, the substituted orunsubstituted alkyl or alkenyl comprises at least 4 carbon atoms(including any carbon atoms in a substituent thereof). In someembodiments, the substituted or unsubstituted alkyl or alkenyl comprisesat least 5 carbon atoms (including any carbon atoms in a substituentthereof).

Examples of hydrocarbon moieties comprising at least one alkyl oralkenyl group include, without limitation, alkyl or alkenyl, optionallysubstituted by hydroxy, cycloalkyl, aryl (e.g., phenyl) and/orheteroaryl (e.g., furanyl); and cycloalkyl, aryl (e.g., phenyl) orheteroaryl (e.g., furanyl) substituted by one or more alkyl or alkenylgroups (e.g., alkyl or alkenyl groups which comprise at least 4 carbonatoms, and optionally at least 5 carbon atoms), and being optionallyfurther substituted (e.g., by one or more hydroxy groups), as theseterms are defined herein.

Without being bound by any particular theory, it is believed that analkyl or alkenyl group in the W hydrocarbon moiety provides the moietywith flexibility (e.g., multiple possible configurations, as a result ofa presence of one or more free-to-rotate bonds, as defined herein) whichenhances interactions thereof (e.g., hydrophobic interactions),especially when the alkyl or alkenyl is of a substantial size (e.g., atleast 4 or 5 carbon atoms, as described herein).

In some embodiments of any of the embodiments described herein wherein Wis alkyl or alkenyl, the alkyl or alkenyl is an unsubstituted alkyl oralkenyl at least 9 carbon atoms in length. In some embodiments, thealkyl or alkenyl is at least 10 carbon atoms in length. In someembodiments, the alkyl or alkenyl is at least 12 carbon atoms in length.

X optionally comprises one or more substituents (in addition to theW-L₁- and -L₂-Y—Z moieties described herein) which comprise ahydrocarbon moiety. Such a substituent is optionally identical to theW-L₁- moiety, according to any of the embodiments described herein. Insome embodiments, X comprises 0 or 1 substituent which is identical tothe W-L₁- moiety.

Y in Formula I is absent or is an aliphatic hydrocarbon moiety beingfrom 1 to 8 atoms in length, which can be substituted or unsubstituted,or alternatively, Y attaches to X and/or to Z to form one or two five-or six-membered cycloalkyl or heteroalicyclic rings.

In embodiments wherein Y forms a heteroalicyclic ring, at least one ringatom in the heteroalicyclic rings is an oxygen atom. In some suchembodiments, one ring atom (and no more) in the heteroalicyclic ring isan oxygen atom, such that the ring is a pyran ring or derivativethereof.

In embodiments wherein Y is absent, L₂ is attached directly to Z, or L₂is absent and Z is —C(═O)OH (a terminal group) attached directly to X.

In some embodiments of any of the embodiments described herein wherein Yis an aliphatic hydrocarbon, the aliphatic hydrocarbon isnon-substituted. In some embodiments, the aliphatic hydrocarbon is asubstituted or unsubstituted linear hydrocarbon. In some embodiments,the aliphatic hydrocarbon is am unsubstituted linear hydrocarbon. Insome embodiments, the aliphatic hydrocarbon is saturated. In someembodiments, the aliphatic hydrocarbon is saturated and non-substituted.In some embodiments, the aliphatic hydrocarbon is a saturated linearhydrocarbon. In some embodiments, the aliphatic hydrocarbon is anunsubstituted and linear saturated hydrocarbon.

In some embodiments of any of the embodiments described herein wherein Yis an aliphatic hydrocarbon, the aliphatic hydrocarbon is from 2 to 7carbon atoms in length. In some embodiments, the aliphatic hydrocarbonis from 3 to 7 carbon atoms in length.

In some embodiments of any of the embodiments described herein wherein Yis an aliphatic hydrocarbon, the aliphatic hydrocarbon is from 1 to 3carbon atoms in length.

Herein, the length of an aliphatic hydrocarbon refers to the number ofcarbon atoms in a hydrocarbon chain separating L₂ and Z, or if Z isabsent, the number of carbon atoms in the longest hydrocarbon chain in Ybeginning with (and including) a carbon atom attached to L₂.

Herein, a “linear” hydrocarbon is a hydrocarbon in which all of thecarbon atoms therein are within a hydrocarbon chain, wherein in thecontext of Y, the hydrocarbon chain begins a carbon atom attached to L₂,and ends in a carbon atom attached to Z, or if Z is absent, ends in aterminal carbon atom (i.e., a carbon atom attached to only one othercarbon atom). The length, in carbon atoms, of a linear hydrocarbonequals the number of carbon atoms in the linear hydrocarbon.

In some embodiments of any of the embodiments described herein wherein Yattaches to X and/or to Z to form one or two five- or six-memberedcycloalkyl or heteroalicyclic rings, Y can be, for example, —CH═CR₇-A-or —CH₂—CHR₈—B—, wherein A and B are each independently absent or anoxygen atom attached to X or to an L₂ group which is —C(═O)— or —C(═O)O—(wherein A or B is attached to the carbon atom of —C(═O)O—), optionally—C(═O)—; and R₇ and R₈ are each independently hydrogen, alkyl and/or a Zmoiety according to any of the respective embodiments described herein.In some such embodiments, the Z moiety is a phenyl substituted by one ormore hydroxy groups, according to any of the respective embodimentsdescribed herein. In some such embodiments, the alkyl represented by R₇and/or R₈ comprises from 1 to 10 carbon atoms, and in some suchembodiments, from 3 to 7 carbon atoms.

Z in Formula I is an optional moiety comprising a polar group, such as,but not limited to, —C(═O)OH and aryl substituted by at least onehydroxy group, or alternatively, Z is absent.

In some embodiments of any of the embodiments described herein wherein Zis aryl, the aryl is hydroxyphenyl or dihydroxyphenyl. In someembodiments, the hydroxyphenyl is 4-hydroxyphenyl and/or thedihydroxyphenyl is 3,4-dihydroxyphenyl.

In some embodiments of any of the embodiments described herein wherein Zis —C(═O)OH, Y is absent or is an aliphatic hydrocarbon moiety from 1 to8 atoms in length, according to any of the respective embodimentsdescribed herein.

In some embodiments of any of the embodiments described herein wherein Zis —C(═O)OH, Y is absent and L₂ is absent or is —C(R₂)(OH)—. In somesuch embodiments, X is cyclic, being, for example, a substituted orunsubstituted bicyclic hydrocarbon moiety, and/or substituted orunsubstituted phenylene, according to any of the respective embodimentsdescribed herein. In some such embodiments, L₂ is absent.

In some embodiments of any of the embodiments described herein wherein Zis —C(═O)OH and Y is an aliphatic hydrocarbon moiety, the aliphatichydrocarbon moiety is from 2 to 7 carbon atoms in length. In some suchembodiments, the aliphatic hydrocarbon moiety is from 3 to 7 carbonatoms in length. In some such embodiments, the aliphatic hydrocarbonmoiety is a saturated hydrocarbon moiety. In some embodiments, thealiphatic hydrocarbon moiety is a linear, unsubstituted hydrocarbonmoiety. In some embodiments, the aliphatic hydrocarbon moiety is alinear and unsubstituted saturated hydrocarbon moiety.

In some embodiments of any of the embodiment described herein wherein Yis an aliphatic hydrocarbon moiety, L₂ is —C(═O)—.

In some embodiments of any of the embodiments described herein wherein Zis —C(═O)OH, and Y is absent or is an aliphatic hydrocarbon moiety, thecompound is a compound having the general formula III:

wherein:

D is CH₂ or —C(═O)—;

the dashed lines each denote a saturated or unsaturated bond;

p is an integer in a range of from 0 to 6;

R₃₀ and R₃₁ are each hydrogen or R₃₀ and R₃₁ to together form anaromatic ring substituted by at least one hydroxy group, wherein whenR₃₀ and R₃₁ are each hydrogen, D is —C(═O)— and each of the dashed linesdenotes an unsaturated bond; and

R₃₂ is an alkyl, alkenyl and/or acyl, each being from 2 to 12 carbonatoms in length.

In some embodiments of any of the embodiments relating to Formula III,the compound is from 16 to 20 carbon atoms in length, e.g., such thatthe sum of the value of p+8, and the length in carbon atoms of R₃₂ is ina range of from 16 to 20. In some embodiments, the compound is from 17to 19 carbon atoms in length. In some embodiments, the compound is 18carbon atoms in length.

In some embodiments of any of the embodiments relating to Formula III,R₃₂ is an unsubstituted alkyl, alkenyl or acyl. In some embodiments R₃₂is an unsubstituted alkyl or acyl.

In some embodiments of any of the embodiments relating to Formula III,R₃₂ is a linear alkyl, alkenyl or acyl. In some such embodiments R₃₂ islinear and unsubstituted alkyl, alkenyl or acyl. Compounds BKT205 andBKT209 are examples of compounds in which R₃₂ is linear andunsubstituted alkyl or acyl.

In some embodiments of any of the embodiments, an aromatic ring formedby R₃₀ and R₃₁ is a phenylene substituted by at least one one hydroxygroup. In some such embodiments, the phenylene is substituted only byone hydroxy group. In some such embodiments, the hydroxy group is at anortho position with respect to the —C(═O)OH group.

In some embodiments of any of the embodiments wherein R₃₀ and R₃₁together form an aromatic ring, D is CH₂. In some such embodiments, thearomatic ring is phenylene, according to any of the respectiveembodiments described herein.

Compound BKT206 is an example of a compound in which D is CH₂, and R₃₀and R₃₁ together form a phenylene substituted only by a hydroxy group atan ortho position with respect to the —C(═O)OH group.

In some embodiments of any of the embodiment described herein wherein Yis an aliphatic hydrocarbon moiety, X is —C≡C—C≡C— or —CR₃═CR₄—CR₅═CR₆—,according to any of the respective embodiments described herein. In someembodiments, X is —CR₃═CR₄—CR₅═CR₆—. In some embodiments, L₂ is —C(═O)—.In some embodiments, L₂ is —C(═O)— and X is —CR₃═CR₄—CR₅═CR₆—.

Returning to the X moiety of Formula I, in some embodiments of any oneof the embodiments described herein relating to Formula I, X is—CR₃═CR₄—CR₅═CR₆—, a substituted or unsubstituted bicyclic hydrocarbonmoiety, and/or a substituted or unsubstituted phenylene, according toany of the respective embodiments described herein.

In some embodiments of any one of the embodiments described hereinrelating to Formula I, X in Formula I is a substituted or unsubstitutedbicyclic hydrocarbon moiety, and/or a substituted or unsubstitutedphenylene, according to any of the respective embodiments describedherein.

In some embodiments of any one of the embodiments described hereinrelating to Formula I, X in Formula I is —CR₃═CR₄—CR₅═CR₆—, and/or asubstituted or unsubstituted phenylene, according to any of therespective embodiments described herein.

In some embodiments of any one of the embodiments described hereinrelating to Formula I, X in Formula I is —C≡C—C≡C— and/or—CR₃═CR₄—CR₅═CR₆—, according to any of the respective embodimentsdescribed herein.

In some embodiments of any of the embodiments described herein wherein Xis —C≡C—C≡C—, L₂ is —C(R₂)(OH)—. In some such embodiments, L₂ is—CH(OH)—.

In some embodiments of any of the embodiments wherein X is —C≡C—C≡C—, L₁is —CR₁(OH)— (e.g., —CH(OH)—). In some such embodiments, L₁ is —CR₁(OH)—and L₂ is —C(R₂)(OH)—. In some such embodiments, L₁ and L₂ are each—CH(OH)—.

In some embodiments of any of the embodiments wherein X is —C≡C—C≡C—, Zis absent.

In some embodiments of any of the embodiments wherein X is —C≡C—C≡C—, Yis an aliphatic hydrocarbon moiety. In some such embodiments Y is anunsaturated aliphatic hydrocarbon moiety (e.g., alkenyl). In some suchembodiments, Y is CH═CH₂.

In some embodiments of any of the embodiments described herein wherein Xis —CR₃═CR₄—CR₅═CR₆—, L₂ is —C(═O)—.

In some embodiments of any of the embodiments wherein X is—CR₃═CR₄—CR₅═CR₆—, L₁ is —C(═O)— or absent. In some such embodiments, L₁is —C(═O)— or absent and L₂ is —C(═O)—.

In some embodiments of any of the embodiments wherein X is—CR₃═CR₄—CR₅═CR₆—, Z is —C(═O)OH.

In some embodiments of any of the embodiments wherein X is—CR₃═CR₄—CR₅═CR₆—, Y is an aliphatic hydrocarbon moiety. In some suchembodiments Y is a saturated hydrocarbon moiety. In some suchembodiments, Y is an unsubstituted saturated hydrocarbon.

In some embodiments of any of the embodiments wherein X is—CR₃═CR₄—CR₅═CR₆—, R₃-R₆ are each hydrogen.

In some embodiments, X is a bicyclic hydrocarbon moiety.

Herein, the phrase “bicyclic hydrocarbon moiety” refers to a hydrocarbonmoiety comprising a pair of carbon atoms covalently bound to each other,wherein the pair of carbon atoms is shared by two fused hydrocarbonrings, each of which may independently be cycloalkyl or aryl.

In some embodiments of any of the embodiments wherein X is a bicyclicmoiety, the bicyclic moiety is an aromatic bicyclic moiety, comprisingtwo aryl rings, for example, a substituted or unsubstitutednaphthalenylene moiety.

In some embodiments of any of the embodiments wherein X is a bicyclicmoiety, the bicyclic moiety is aliphatic (i.e., alicyclic), that is,both hydrocarbon rings are cycloalkyl.

In some embodiments of any of the embodiments wherein X is a bicyclicmoiety, at least one of the two fused hydrocarbon rings is asix-membered ring.

In some embodiments, the two fused hydrocarbon rings are bothsix-membered ring. That is, X is a substituted or unsubstituted decalinmoiety, or an unsaturated derivative thereof (a dehydrogenated decalinmoiety).

In some embodiments of any of the embodiments wherein X is a bicyclicmoiety, the bicyclic moiety is unsaturated, that is, at least two ringcarbons are attached via an unsaturated bond. In some such embodiments,the bicyclic moiety is a substituted or unsubstituted dehyrogenateddecalin. In some such embodiments, the dehydrogenated decalin is asubstituted or unsubstituted 1,2,3,4,5,6,7,8-octahydronaphthalenemoiety, wherein the unsaturated carbon-carbon bond is the bond shared bythe two fused rings.

In some embodiments of any of the embodiments wherein X is a substitutedbicyclic moiety, the substituent(s) of the bicyclic moiety is alkyl. Insome embodiments, the alkyl is C₁₋₄ alkyl. In some embodiments, thesubstituent(s) of the bicyclic moiety is methyl.

In some embodiments of any of the embodiments wherein X is a substitutedbicyclic moiety, L₂ or Z is attached to a carbon atom in X which isshared by two fused rings or to a carbon atom in X which is directlyattached to a carbon atom shared by two fused rings.

In some embodiments of any of the embodiments wherein X is a substitutedbicyclic moiety, W or L₁ is attached to a carbon atom in X which isshared by two fused rings or to a carbon atom in X which is directlyattached to a carbon atom shared by two fused rings.

Without being bound by any particular theory, it is believed that abicyclic structure can provide rigidity to the X moiety, particularly tothe portion of the bicyclic structure represented by the carbon atomsshared by two fused rings and carbon atoms attached thereto.

In some embodiments, X is a phenylene moiety.

Herein, the term “phenylene” refers to a substituted or unsubstitutedsix-membered aromatic carbon ring having at least two valence bonds(e.g., one bond to L₁ or W, and one bond to L₂ or Z, in the context ofthe X moiety of Formula I), and optionally more than two valence bonds(e.g., one bond to L₁ or W, and one bond to L₂, and one bond to Y, inthe context of the X moiety of Formula I).

In some embodiments of any of the embodiments wherein X is a substitutedphenylene, the phenylene is substituted by at least one substituent,optionally comprising one or more hydroxy and/or a saccharide moieties.In some such embodiments, X is substituted by one or two suchsubstituents. In some embodiments, the saccharide moiety is amonosaccharide moiety. In some embodiments, the saccharide moiety is ahexose moiety. In some embodiments, the saccharide moiety is a glucosemoiety.

The saccharide moiety may optionally be bound to the phenylene ring, forexample, via a carbon atom in the saccharide moiety (e.g., via aC-glycosidic bond), via an oxygen atom in the saccharide moiety (e.g.,via an O-glycosidic bond), via a sulfur atom in the saccharide moiety(e.g., via an S-glycosidic bond) or via a nitrogen atom in thesaccharide moiety (e.g., via an N-glycosidic bond). In some embodimentsof any of the respective embodiments described herein, the phenylenering is attached to an oxygen atom of the saccharide moiety. In someembodiments, the saccharide moiety and phenylene are attached via anO-glycosidic bond.

Herein, the term “glycosidic bond” refers to a bond with an anomericcarbon of a saccharide moiety, optionally via a heteroatom (e.g., oxygenatom) attached to the anomeric carbon.

In some embodiments of any of the embodiments wherein X is a substitutedphenylene, the substitution is in a form of a cycloalkyl,heteroalicyclic, aryl or heteroaryl ring fused to the phenylene, eachbeing substituted or unsubstituted. It is to be understood that such aring is not a ring formed by a Y moiety described herein (as the Ymoiety is not considered herein to be a substituent). In some suchembodiments, the ring is a heteroalicyclic or heteroaryl ring(optionally unsubstituted) comprising an oxygen atom attached to acarbon atom of the phenylene ring. In some such embodiments, the ring isa furan ring (optionally unsubstituted) fused to the phenylene to form abenzofuran moiety. Compound BKT208 is an example of a compound in whichX is such a benzofuran moiety.

In some embodiments of any of the embodiments wherein X is phenylene, Yattaches to X to form a six-membered ring wherein at least one ring atomis an oxygen atom, according to any of the respective embodimentsdescribed herein. In some such embodiments, no more than one ring atomis an oxygen atom. In some such embodiments, the ring is substituted orunsubstituted pyrone ring (e.g., a 2-pyrone ring or 4-pyrone ringwherein L₂ is —C(═O)— and Y is —CH═CHR₇—O—, or a 2-pyrone wherein L₂ is—C(═O)O— and Y is —CH═CHR₇—) or a dihydropyrone ring (e.g., wherein L₂is —C(═O)— and Y is —CH₂═CHR₈—O—, or L₂ is —C(═O)O— and Y is—CH₂═CHR₈—).

According to an aspect of embodiments of the invention, there isprovided a compound having the general formula II:

for use in modulating a biological activity of a chemokine, for use ininhibiting SDF-1 and/or CXCR4 and/or in treating a cancer, aninflammation and/or a non-cancerous hyperproliferative disorder,according to any of the respective embodiments described herein,

wherein:

each of the dashed lines independently denotes a saturated orunsaturated bond;

J is —O— (when the dashed line between J and K denotes a saturated bond,and J is thus divalent) or —CR₁₄═(when the dashed line between J and Kdenotes an unsaturated bond, and J is trivalent);

K is —CR₁₅═(when K is trivalent), or is —CR₁₆R₁₇— or —C(═O)— (when K isdivalent);

M is —CR₁₈═(when M is trivalent), or is —CR₁₉R₂₀— or —C(═O)— (when K isdivalent);

Q is —C(═O)— (when the dashed line between Q and M denotes a saturatedbond, and Q is thus divalent) or —CR₂₁═(when the dashed line between Qand M denotes an unsaturated bond, and Q is thus trivalent);

R₁₀, R₁₂ and R₁₃ are each independently hydrogen, hydroxy, alkyl,alkenyl, alkynyl, cycloalkyl, alkoxy and/or aryloxy;

R₁₁ is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl or heteroalicyclic,or alternatively, R₁₁ and R₁₂ together form a five- or six-memberedheteroaryl or heteroalicyclic ring;

R₁₄-R₂₁ are each independently hydrogen, alkyl, alkenyl, alkynyl,cycloalkyl and/or aryl;

and at least one of K and Q is —C(═O)—, and at least one of J and M is—O—.

Whether the dashed line denotes a saturated or unsaturated bond will beclear to the skilled person based on the valences of J, K, M and Q. Forexample, the dashed line linking J and K denotes an unsaturated bondonly if J is CR₁₄ and K is CR₁₅; the dashed line linking K and M denotesan unsaturated bond only if K is CR₁₅ and M is CR₁₈; and the dashed linelinking M and Q denotes an unsaturated bond only if M is CR₁₈ and Q isCR₂₁.

In some embodiments of any of the embodiments described herein withrespect to Formula II, the compound of Formula II is a compound ofFormula I, as described herein, wherein X is the phenylene ring attachedto R₁₀, R₁₂ and R₁₃; at least one of R₁₀-R₁₃ is a W moiety (as definedaccording to any of the respective embodiments described herein); Q is—C(═O)—, corresponding to L₂, and M, K and J together correspond to Y(optionally further including Z), or alternatively, J and K together are—C(═O)O—, corresponding to L₂, and M and Q together correspond to Y(optionally further including Z).

In some embodiments of any of the embodiments described herein withrespect to Formula II, no more than one of K and Q is —C(═O)—.

In some embodiments of any of the embodiments described herein withrespect to Formula II, no more than one of J and M is —O—. In some suchembodiments, one of K and Q is —C(═O)— and one of J and M is —O—, e.g.,such that the ring formed by J, K, M and Q is a pyrone or dihydropyronering.

In some embodiments of any of the embodiments described herein withrespect to Formula II, R₁₇-R₂₁ are each hydrogen.

In some embodiments of any of the embodiments described herein withrespect to Formula II, R₁₀ and R₁₂ are each independently hydrogen,alkyl and/or alkenyl. In some embodiments, the alkyl and alkenyl areunsubstituted. In some embodiments, the alkyl and alkenyl have up to 6carbon atoms (i.e., the alkyl has from 1 to 6 carbon atoms and thealkenyl has from 2 to 6 carbon atoms). In some embodiments, the alkyland alkenyl have from 4 to 6 carbon atoms. Isoprenyl (3-methyl2-butenyl) is an exemplary alkenyl group. Compound BKT201 is an exampleof a compound wherein R₁₀ and R₁₂ are hydrogen and/or isoprenyl.

In some embodiments of any of the embodiments described herein withrespect to Formula II, R₁₀ and R₁₂ are alkyl and/or alkenyl, having upto 3 carbon atoms (i.e., the alkyl has from 1 to 3 carbon atoms and thealkenyl has 2 or 3 carbon atoms). In some such embodiments, the alkyl ismethyl. Compound BKT204 is an example of a compound, in which R₁₀ andR₁₂ are each methyl.

In some embodiments of any of the embodiments described herein withrespect to Formula II, J is —O— or —CH═. In some such embodiments, J is—O—.

In some embodiments of any of the embodiments described herein withrespect to Formula II, J is —O— and K is —CR₁₆R₁₇—.

In some embodiments of any of the embodiments described herein withrespect to Formula II, R₁₆ is alkyl or phenyl, each being substituted orunsubstituted. In some embodiments, the alkyl is unsubstituted (e.g.,pentyl). In some embodiments, the phenyl is substituted by one or morehydroxy groups. In some such embodiments, the phenyl is hydroxyphenyl(e.g., 3, hydroxyphenyl) or dihydroxyphenyl (e.g., 3,4-dihydroxyphenyl).

In some embodiments of any of the embodiments described herein withrespect to Formula II, Q is —C(═O)—. In some such embodiments, J is —O—or —CH═, optionally —O—. In some such embodiments, K is —CR₁₆R₁₇—wherein R₁₆ is alkyl or phenyl, each being substituted or unsubstituted.Compounds BKT201, BKT204 and BKT300 are examples of compounds in which Qis —C(═O)—.

In some embodiments of any of the embodiments described in which Q is—C(═O)—, R₁₁ is hydrogen or glucosyl. Compounds BKT204 and BKT300 areexamples of compounds in which Q is —C(═O)— and R₁₁ is hydrogen.Compound BKT201 is an example of a compound in which R₁₁ is glucosyl.

In some embodiments of any of the embodiments described herein withrespect to Formula II, M is —CH₂—. In some such embodiments, Q is—C(═O)—. In some such embodiments, J is —O—. In some such embodiments, Qis —C(═O)— and J is —O—.

In some embodiments of any of the embodiments described herein withrespect to Formula II, R₁₃ is —OH, alkoxy or aryloxy (optionally —OH).In some such embodiments, Q is —C(═O)— and J is —O—. In some suchembodiments, M is —CH₂—. In some such embodiments, Q is —C(═O)—, J is—O— and M is —CH₂—. Compounds BKT201 and BKT204 are examples ofcompounds in which M is CH₂, Q is —C(═O)— and R₁₃ is —OH.

In some embodiments of any of the embodiments described herein withrespect to Formula II, M is —O—. In some such embodiments, Q is —C(═O)—.In some such embodiments, J is —CH═. In some such embodiments, Q is—C(═O)— and J is —CH═. Compound BKT300 is an example of a compound inwhich M is —O—, Q is —C(═O)— and J is —CH═.

In some embodiments of any of the embodiments described herein withrespect to Formula II, K is —C(═O)—.

In some embodiments of any of the embodiments described herein withrespect to Formula II, M and Q are linked via an unsaturated bond. Insome such embodiments, M and Q are each —CH═. In some such embodiments,K is —C(═O)—. In some such embodiments, K is —C(═O)— and M and Q areeach —CH═.

In some embodiments of any of the embodiments described herein withrespect to Formula II, R₁₃ is hydrogen. In some such embodiments, K is—C(═O)— and J is —O.

In some such embodiments, M and Q are each is —CH═. In some suchembodiments, K is —C(═O)—, J is —O— and M and Q are each is —CH═.Compounds BKT202, BKT208 and BKT215 are examples of compounds in whichR₁₃ is hydrogen, K is —C(═O)—, J is —O— and M and Q are each —CH═.

In some embodiments of any of the embodiments described herein wherein Kis —C(═O)— and J is —O—, R₁₀ and/or R₁₂ is hydrogen. In some suchembodiments, both R₁₀ and R₁₂ are hydrogen.

In some embodiments of any of the embodiments described herein wherein Kis —C(═O)— and J is —O—, R₁₁ is substituted or unsubstituted alkyl,alkenyl or cycloalkyl, each having from 4 to 20 carbon atoms, optionallyfrom 5 to 20 carbon atoms. In some such embodiments, R₁₀ and/or R₁₂ ishydrogen. In some such embodiments, both R₁₀ and R₁₂ are hydrogen. Insome such embodiments, R₁₁ is substituted or unsubstituted alkyl oralkenyl (e.g., as in Compounds BKT202 and BKT215).

In some embodiments of any of the embodiments described herein wherein Kis —C(═O)— and J is —O—, Ru and R₁₂ together form a furan ring. CompoundBKT208 is an example of such a compound.

According to an aspect of some embodiments of the present invention,there is provided a compound which is an anthracene dione derivativehaving a general formula which is Formula IV (which comprises twoanthracene moieties):

or Formula V (which comprises one anthracene moiety):

wherein:

R₄₀-R₅₁ and R₆₁-R₆₈ are each hydrogen, hydroxy, alkyl, C-carboxy and/ora saccharide moiety;

R₅₂-R₅₅ are hydrogen, or alternatively, R₅₂ and R₅₃, and/or R₅₄ and R₅₅,are together a covalent bond which forms a six-membered carbon ring; and

the dashed line denotes a saturated or unsaturated bond.

It is to be appreciated that R₆₁-R₆₃ and R₆₆R₆₈ in Formula V correspondrespectively to R₄₃-R₄₅ and R₄₀-R₄₂ and/or to R₄₉-R₅₁ and R₄₆-R₄₈ inFormula IV.

In some embodiments of any of the embodiments relating to Formulas IVand V, R₄₀-R₅₁ and R₆₁-R₆₈ are each hydrogen, hydroxy, alkyl and/oralkylcarboxy.

In some embodiments of any of the embodiments relating to Formulas IVand V, the alkyl is methyl and/or the C-carboxy is methylcarboxy (i.e.,CH₃—O—C(═O)—).

In some embodiments of any of the embodiments relating to Formulas IVand V, R₄₁, R₄₄, R₄₇, R₅₀, R₆₁-R₆₃ and R₆₈ are each hydrogen.

In some embodiments of any of the embodiments relating to Formula IV,the dashed line denotes an unsaturated bond, R₅₂ and R₅₃ are together acovalent bond which forms a six-membered aromatic carbon ring, and R₅₄and R₅₅ are together a covalent bond which forms a six-membered aromaticcarbon ring. Compound BKT203 is an example of such a compound.

In some embodiments of any of the embodiments relating to Formula IV,the dashed line denotes a saturated bond, and R₅₂-R₅₅ are each hydrogen.Compound BKT210 is an example of such a compound.

According to an aspect of some embodiments of the present invention,there are provided compounds, which are collectively represented byFormula VI:

wherein:

A′ is selected from the group consisting of —C(═O)—O—; —NR₇₁—C(═O)—;—C(R₇₂)—N═; —C(R₇₂)═N—; and —C(R₇₃)═C(R₇₄)—;

B is selected from the group consisting of NR₇₅—C(═O)—; C(R₇₆)—N═;—C(R₇₆)═N—; —C(R₇₇)—C(═O)—; and —C(R₇₇)═C(R₇₈)—;

Y′₁ and Y′₂ are each independently selected from C-Q′, C—R₇₉ and N,provided

that at least one of Y′₁ and Y′₂ is C-Q′;

Y′₃ is selected from N and C—R₈₀;

Y′₄ is selected from N and C—R₈₁;

R₇₁ and R₇₅ are each independently hydrogen or alkyl;

R₇₂-R₇₄ are each independently selected from hydrogen, alkyl, alkyne,hydroxy, amine, alkoxy, aryloxy, thioalkoxy, thioaryloxy, carbonyl andcarboxylate;

R₇₆-R₇₈ are each independently selected from hydrogen, alkyl, hydroxy,amine, alkoxy, aryloxy, thioaryloxy, carboxylate, cinnamic acid, acyl,S(OH)₃ and S—O—O—OH;

R₇₉ is hydrogen or cyano;

R₈₀ and R₈₁ are each independently selected from hydrogen, hydroxy,halo, alkoxy, thioalkoxy, aryloxy, thioaryloxy, thiol, amine, and SO₃H;and

Q′ is

wherein:

Z′ is selected from 0, NH, C(═O), S, CH₂ and S(═O);

X′₁ is C(R₈₂) or N;

X′₂ is C(R₈₃) or N;

X′₃ is C(R₈₄) or N;

X′₄ is C(R₈₅) or N; and

X′₅ is C(R₈₆) or N,

provided that at least two of X′₁-X′₅ are not N,

R₈₂ is selected from hydrogen, alkyl and halo; and

R₈₃-R₈₆ are each independently selected from hydrogen, halo, alkyl,amine,

alkoxy, aryloxy, carboxy and hydroxy;

or, alternatively, two of R₈₂-R₈₆ are joined together to form an aryl orheteroaryl.

In some of any of the embodiments described herein for Formula VI,R₈₃-R₈₆ are each independently selected from hydrogen, halo, alkyl,amine, alkoxy, aryloxy and hydroxy.

In some of any of the embodiments described herein for Formula VI, A′ isselected from the group consisting of —C(R₇₂)—N═; —C(R₇₂)═N—; and—C(R₇₃)═C(R₇₄)—; and

B′ is selected from the group consisting of —C(R₇₆)—N═; —C(R₇₆)═N—; andC(R₇₇)═C(R₇₈).

In some of any of the embodiments described herein for Formula VI, Y′₁is C-Q′.

In some of any of the embodiments described herein for Formula VI, A′ isselected from the group consisting of —C(R₇₂)—N═; —C(R₇₂)═N—; and—C(R₇₃)═C(R₇₄)—; B′ is selected from the group consisting of —C(R₇₆)—N═;—C(R₇₆)═N—; and —C(R₇₇)═C(R₇₈)—; and Y′₁ is C-Q′.

In some of these embodiments, Y′₂ is C—R₇₉; Y′₃ is C—R₈₀; and Y′₄ isselected from N and C—R₈₁.

Exemplary such compounds are presented in Table 4 herein below.

In some of any of the embodiments described herein for Formula VI, Y′₁is C-Q′, and Z′ is O, NH, C(═O), S, CH₂ or S(═O).

In some of any of the embodiments described herein for Formula VI, A′ is—C(R₇₃)═C(R₇₄)— and B′ is —C(R₇₇)═C(R₇₈)—, such that the bicyclicskeleton presented in Formula VI is of a naphthalene.

In some of these embodiments, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃ is C—R₈₀;and Y′₄ is C—R₈₁.

In some of these embodiments, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃ is C—R₈₀;and Y′₄ is N.

In some of any of the embodiments described herein for Formula VI, A′ is═C(R₇₂)—N═; and B′ is —C(R₇₆)—N═, such that the bicyclic skeletonpresented in Formula VI is of a quinazoline. Such a skeleton canalternatively be presented as A′ being —C(R₇₂)═N—; and B′ being—C(R₇₆)═N—.

In some of these embodiments, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃ is C—R₈₀;and Y′₄ is C—R₈₁.

In some of any of the embodiments described herein for Formula VI, A′ is—C(R₇₃)═C(R₇₄)—; B is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃ isC—R₈₀; Y′₄ is C—R₈₁, and Z′ is NH.

In some of these embodiments, one or more of R₇₇, R₈₀ and R₈₁ issulfonate (—S(═O)₂—OH).

In some of these embodiments, R₈₀ and R₈₁ is sulfonate (—S(═O)₂—OH), andR₇₇ is —S(OH)₃.

In some of any of the embodiments of Formula VI, in which A′ is—C(R₇₃)═C(R₇₄)—; B′ is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; Y′₄ is C—R₈₁, and Z′ is NH, R₇₃ is OH.

In some of any of the embodiments of Formula VI, in which A′ is—C(R₇₃)═C(R₇₄)—; B′ is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; Y′₄ is C—R₈₁, and Z′ is NH, Q′ is such that X′₁, X′₃ and X′₅are each N, and X′₂ and X′₄ are each C—R₇₃ and C—R₇₅, respectively.

In some of these embodiments, R₈₃ and R₈₅ are each independentlyselected from halo, alkyl, and amine. In some of these embodiments, theamine is a secondary amine, such that it is substituted by e.g., asubstituted or unsubstituted alkyl or aryl.

In some of any of the embodiments described herein for Formula VI, A′ is—C(R₇₃)═C(R₇₄)—; B′ is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; Y′₄ is C—R₈₁, and Z′ is —C(═O) or —S(═O).

In some of these embodiments, Z′ is C═O, and R₇₃ is OH.

In some of these embodiments, Z′ is C═O, R₇₃ is OH, and R₇₈ is carbonyl,—C(═O)—R, with R being, for example, phenyl.

In some of these embodiments, Q′ is phenyl, for example, unsubstitutedphenyl, (such that X₇₁—X₇₅ are each C—H), or a substituted phenyl, inwhich one or more of R₈₂-R₈₆ is other than H.

In some of the embodiments in which A′ is —C(R₇₃)═C(R₇₄)—; B′ is—C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y₂ is C—R₇₉; Y′₃ is C—R₈₀; Y′₄ is C—R₈₁,and Z′ is —C(═O), R₇₃ is OH, Q′ is phenyl, and one or more of R₇₇, R₇₈,R₈₀ and R₈₁ is sulfonate.

In some of any of the embodiments described herein for Formula VI, A′ is—C(R₇₃)═C(R₇₄)—; B′ is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; Y′₄ is C—R₈₁, and Z′ is —S(═O).

In some of these embodiments, R₇₃ is carboxy.

In some of any of these embodiments, Q′ is phenyl (substituted orunsubstituted). In some of these embodiments, the phenyl is substitutedby one or more halo substituents.

In some of any of these embodiments, one or more of R₇₄, R₇₇ and R₇₈ isselected from hydroxy and alkoxy.

In some of any of the embodiments described herein for Formula VI, A′ is—C(R₇₃)═C(R₇₄)—; B′ is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; Y′₄ is C—R₈₁, and Z′ is CH₂.

In some of any of these embodiments, one or more of R₇₄, R₇₇, R₇₈, R₈₀and R₈₁ is selected from hydroxy and alkoxy.

In some of any of these embodiments, R₇₃ is alkyne, which can besubstituted or unsubstituted.

In some of any of these embodiments, Q′ is such that 2 of X′₁-X′₅ arenitrogen atoms.

In some of any of these embodiments, Q′ is phenyl.

In some of any of the embodiments described herein for Formula VI, A′ is—C(R₇₃)═C(R₇₄)—, B′ is —C(R₇₇)═C(R₇₈)—, Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; Y′₄ is C—R₈₁, and Z′ is O or S.

In some of any of the embodiments described herein for Formula VI, A′ is═C(R₇₂)—N═; B′ is —C(R₇₆)—N═; Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃ is C—R₈₀;and Y′₄ is C—R₈₁.

In some of these embodiments, R₇₂ and R₇₆ are each an amine.

In some of these embodiments, R₇₉ is cyano (nitrile).

In some of these embodiments, R₈₁ is halo (e.g., fluoro or chloro).

In some of these embodiments, R₈₀ is a group that comprises an aryl(e.g., phenyl). In some embodiments, R₈₀ is anilino (—NH-phenyl), oraryloxy (e.g., —O— phenyl) or thioaryloxy.

In some of any of the embodiments described herein for Formula VI, inwhich A′ is ═C(R₇₂)—N═; B′ is —C(R₇₆)—N═(or, alternatively, A′ is—C(R₇₂)═N—; and B′ is —C(R₇₆)═N—); Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃ isC—R₈₀; and Y′₄ is C—R₈₁, Z′ is S, O or NH.

In some of any of the embodiments described herein for Formula VI, inwhich A′ is ═C(R₇₂)—N═; B′ is —C(R₇₆)—N═; Y′₁ is C-Q′, Y′₂ is C—R₇₉; Y′₃is C—R₈₀; and Y′₄ is C—R₈₁, Q′ is phenyl, and in some embodiments, it isan unsubstituted phenyl (such that X′₁-X′₅ are each C—H).

In some of any of the embodiments described herein, the compoundfeatures an additional aromatic moiety (aryl or heteroaryl, preferablyaryl, as defined herein). In some of these embodiments, the aromaticmoiety is or forms a part of one or more of R₇₂-R₇₄, R₇₆-R₈₁, andR₈₂-R₈₆, when feasible.

In some of these embodiments, the aromatic moiety is in a form of anamine substituted by an aryl, an aryloxy, a thioaryloxy, and alkaryl, oras carboxyaryl.

In some of any of the embodiments described herein, the compounds arecollectively represented by Formula VIA:

wherein:

A′ is selected from the group consisting of —C(═O)—O— and —NR₇₁—C(═O)—;

B′ is selected from the group consisting of —NR₇₅—C(═O)—; —C(R₇₇)—C(═O)—and —C(R₇₇)═C(R₇₈)—;

Y′₁ and Y′₂ are each independently selected from C-Q′, CH and N,provided that at least one of Y′₁ and Y′₂ is C-Q′;

R₇₁ and R₇₅ are each independently hydrogen or alkyl; and

R₇₇ and R₇₈ are each independently hydrogen and alkyl,

at least one of R₇₁, R₇₇ and R₇₈ is an alkyl being at least 4 atoms inlength;

R₈₀ and R₈₁ are each independently selected from hydrogen and hydroxyl,at least one of R₈₀ and R₈₁ being hydroxy; and

Q′ is

wherein:

Z′ is O;

X′₁ is C(R₈₂);

X′₂ is C(R₈₃) or N;

X′₃ is C(R₈₄);

X′₄ is C(R₈₅) or N; and

X′₅ is C(R₈₆) or N,

R₈₂ is selected from hydrogen and alkyl; and

R₈₃-R₈₆ are each independently selected from hydrogen, alkoxy, aryloxyand hydroxyl, at least one of R₈₃-R₈₆ being hydroxyl.

In some of any of the embodiments of Formula VIA, R₈₂ is an alkyl beingat least 4 carbon atoms in length.

In some of any of these embodiments, A′ is —C(═O)—O—; and B′ is—C(R₇₇)═C(R₇₈)—. In some of these embodiments, R₇₇ is an alkyl being atlast 4 carbon atoms in length.

In some embodiments of Formula VIA, R₈₂ is an alkyl being at least 4carbon atoms in length, and R₇₇ is an alkyl being at last 4 carbon atomsin length.

In some of any of the embodiments of Formula VIA, Y′₁ is C-Q′ and Y′₂ isC—R₇₉.

In some of these embodiments, R₈₀ is OH or alkoxy.

In some of any of the embodiments of Formula VIA, A′ is —NR₇₁—C(═O)—;and B′ is —NR₇₅—C(═O)—.

In some of these embodiments, R₇₁ is an alkyl being 4 carbon atoms inlength.

In some of any of the embodiments of Formula VIA, A′ is —NR₇₁—C(═O)—;and B′ is —C(R₇₇)—C(═O)—.

In some of these embodiments, R₇₇ is an alkyl being 4 carbon atoms inlength.

In some of these embodiments, Y′₁ is C-Q′ and Y′₂ is C—R₇₉.

In some of these embodiments, R₈₀ is OH or alkoxy.

In some of these embodiments each of X′₁-X′₅ is other than N, such thatQ′ is an aryl, more specifically phenyl.

In some of any of the embodiments of Formula VIA, A′ is —NR₇₁—C(═O)—;and B′ is —NR₇₅—C(═O)—; Y′₁ is N and Y′₂ is C-Q′.

In some of these embodiments, R₇₁ is an alkyl being 4 carbon atoms inlength.

In some of these embodiments, R₈₁ is OH or alkoxy.

In some of any of the embodiments described herein for Formula VIA, eachof X′₁-X′₅ is other than N, such that Q′ is an aryl, more specificallyphenyl.

In some of any of the embodiments described herein for Formula VIA, atleast one of X′₂, X′₄ or X′₅ is N, such that Q′ is a heteroaryl, morespecifically, a nitrogen-containing heteroaryl.

In some of any of the embodiments described herein for Formula VIA, atleast two of X′₂, X′₄ or X′₅ are N.

The nitrogen-containing heteroaryl can be, for example, pyridine, in acase of one nitrogen atom; pyridazine, pyrimidine or pyrazine, in a caseof 2 nitrogen atoms; 1,2,4-triazine, or 1,2,3-triazine, in a case of 3nitrogen atoms; or 1,2,3,4-tetrazine, 1,2,3,5-tetrazine, or1,2,4,5-tetrazine, in a case of 4 nitrogen atoms, each can besubstituted or unsubstituted, preferably substituted, as defined forR₈₂-R₈₆ in Formula VIA.

In some of any of the embodiments described herein for Formula VIA, R₈₄is hydroxyl.

Exemplary compounds represented by Formula VIA as described herein arepresented in Table 5 hereinafter.

Method of Identifying Small Molecules:

According to an aspect of some embodiments of the present invention,there is provided a method of identifying compounds which are usable inany of the methods and uses described herein. The method is generallyeffected by determining a pharmacophoric binding site of at least onekinase in which a small molecule is capable of interacting with at leasttwo amino acid residues which are required for the activity of thekinase; determining a topology of BKT300 which exhibits interaction withthe at least two amino acid residues in the pharmacophoric binding site;and screening a computer-readable database so as to identify a compoundfeaturing a topology similar to the topology of BKT300, wherein acompound featuring the topology is identified as usable in inhibiting atleast one kinase and/or in treating a disease or disorder associatedwith an activity of at least one kinase, and/or in modulating abiological activity of a chemokine and/or on treating cancer.

Herein throughout, the term “topology” means a spatial arrangement (2Dor 3D arrangement) of the chemical groups composing the compound. Forexample, BKT300 is comprised of a bicyclic heteroaromatic moiety whichis linked to an aromatic moiety via an oxygen heteroatom, and whichfurther comprises 2 flanking alkyl groups. In the computational dockingassays performed, it was determined that BKT300 is arranged in thebinding site of kinases in a certain conformation, namely, theheteroaromatic, aromatic and flanking alkyl groups are spatiallyarranged in that certain conformation. This conformation is referred toherein as a “topology” similar to the topology of BKT300.

According to some embodiments of this aspect of the present invention,the method further comprises computationally docking the compound withinthe pharmacophoric binding site, wherein a compound that features in thedocking a spatial arrangement that allows it to interact with the atleast two amino acid residues in the binding site is determined asusable in inhibiting the at least one kinase and/or in treating thedisease or disorder associated with an activity of the at least onekinase.

According to some embodiments of this aspect of the present invention,the pharmacophoric binding site is constructed upon aligning structuresof at least two kinases.

According to some embodiments of this aspect of the present invention,the kinase is selected from MELK and MAPK4K.

According to some embodiments of this aspect of the present invention,the at least two amino acid residues within the pharmacophoric bindingsite comprise Lys40 and Asp150.

According to some embodiments of this aspect of the present invention,the compound is usable in modulating a biological activity of achemokine, and the method further comprises determining a biologicalactivity of the chemokine in the presence of the compound featuring thetopology, for example, as described herein in the Examples section thatfollows.

According to some embodiments of this aspect of the present invention,the compound is usable in treating cancer, and the method furthercomprises determining an anti-cancer activity of the compound featuringthe topology, for example, as described herein in the Examples sectionthat follows. According to some of these embodiments, determining theanti-cancer activity is performed using compounds featuring thetopology, which are further determined as modulators of a biologicalactivity of a chemokine.

According to some embodiments of this aspect of the present invention,the compound is usable in treating inflammation, and the method furthercomprises determining an anti-inflammatory activity of the compoundfeaturing the topology, using methods well known in the art. Accordingto some of these embodiments, determining the anti-inflammatory activityis performed using compounds featuring the topology, which are furtherdetermined as modulators of a biological activity of a chemokine.

According to some embodiments of this aspect of the present invention,the compound is usable in inducing cell death, and the method furthercomprises determining an cell-killing activity of the compound featuringthe topology, for example, as described herein in the Examples sectionthat follows. According to some of these embodiments, determining thecell-killing activity is performed using compounds featuring thetopology, which are further determined as modulators of a biologicalactivity of a chemokine.

As exemplified herein, the method of identifying compounds as describedherein was used to identify compounds having Formula VI.

Therapeutic Applications:

According some embodiments, a small molecule compound of Formula I, II,III, IV, V, VI, VIA and/or VIB, and/or a compound represented in Table2, as described herein in any of the respective embodiments, and anycombination thereof, is capable of, or is useful in, inhibiting abiological activity of a kinase, and/or inhibiting cancer cells, and/orkilling cancer cells, and/or inducing apoptosis, and/or inducing growtharrest, and/or inhibiting chemokine-dependent cell migration, and/ormodulating a biological activity of a chemokine (e.g., cell migration),and/or treating diseases and disorders associated with kinase activityand/or cell migration, such as cancer and inflammatory diseases anddisorders; and/or treating proliferative diseases or disorders (whereinducing apoptosis and/or growth arrest is desirable), as describedherein.

For example, the compounds of Formula VI, according to some embodimentdescribed herein, and any combination thereof, can be regarded asstructural analogs of BKT300, an exemplary compound which is shownherein to act as a chemokine-binding compound, by modulating abiological activity of chemokines, as an inhibitor ofchemokine-dependent cell migration, as an inhibitor of cancer cells(e.g., as inhibitor of cancer cells growth and/or as inducing apoptosisand/or as inhibitor of cancer cells migration), and/or as a kinaseinhibitor.

As inflammation and cancer are typically governed by cell migration(e.g., infiltration, metastasis) and kinase activity, which is oftenassociated with cell proliferation, such conditions are contemplated fortreatment using the compounds of some embodiments of the invention.

Proliferative diseases and disorders as described herein, includingmedical conditions other than cancer (also referred to herein as“non-cancerous hyperproliferative diseases”), are also contemplated fortreatment using the compounds of some embodiments of the invention, dueto the apoptosis-inducing effect of the compounds.

Without being bound by any particular theory, it is believed that thecompounds described herein are particularly useful as anti-cancer agentsby inducing cancer cell death, by affecting chemokine-dependent cancercell migration (e.g., by inhibiting metastasis) and/or angiogenesis,and/or by inhibiting kinase activity (e.g., pro-proliferation kinaseactivity) and/or by inducing apoptosis of cancer cells and/or byinducing growth arrest of cancer cells; and/or as anti-inflammatoryagents by affecting chemokine-dependent immune cell migration (e.g.,immune cell infiltration) and/or by inhibiting kinase activity (e.g.,pro-inflammatory kinase activity), as described in further detail hereinbelow.

In some of any of the embodiments described herein, a small moleculecompound as described herein in any of the respective embodiments, iscapable of, or usable in, inducing death of pathogenic cells (e.g.,cancer cells or immune cells or hyper proliferative cells).

In some of any of the embodiments described herein, a small moleculecompound as described herein in any of the respective embodiments, iscapable of, or usable in, inducing cell death of pathogenic cells.

As used herein, the term “apoptosis” refers to an intrinsic cellself-destruction or suicide program. In response to a triggeringstimulus, cells undergo a cascade of events including cell shrinkage,blebbing of cell membranes and chromatic condensation and fragmentation.These events culminate in cell conversion to clusters of membrane-boundparticles (apoptotic bodies), which are thereafter engulfed bymacrophages.

Methods of monitoring cellular changes induced by the compounds areknown in the art and include for example, the MTT test which is based onthe selective ability of living cells to reduce the yellow salt MTT(3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (Sigma,Aldrich St Louis, Mo., USA) to a purple-blue insoluble formazanprecipitate; the BrDu assay [Cell Proliferation ELISA BrdU colorimetrickit (Roche, Mannheim, Germany]; the TUNEL assay [Roche, Mannheim,Germany]; the Annexin V assay [ApoAlert® Annexin V Apoptosis Kit(Clontech Laboratories, Inc., CA, USA)]; the Senescenceassociated-β-galactosidase assay (Dimri GP, Lee X, et al. 1995. Abiomarker that identifies senescent human cells in culture and in agingskin in vivo. Proc Natl Acad Sci USA 92:9363-9367); 7-ADD viabilitystaining (available from MDsystems), caspase-3 assay (available fromMDsystems) as well as various RNA and protein detection methods (whichdetect level of expression and/or activity) which are further describedhereinabove.

In some of any of the embodiments described herein, a small moleculecompound as described herein in any of the respective embodiments, thecellular change is apoptosis such as via cleavage of caspase-3.

In some of any of the embodiments described herein, a small moleculecompound as described herein in any of the respective embodiments, iscapable of, or usable in, inducing apoptosis via cleavage of caspase-3.

In some of any of the embodiments described herein, a small moleculecompound as described herein in any of the respective embodiments, iscapable of, or usable in, inducing growth arrest of cells, and in someembodiments, the arrest is at the G2M phase of the cell cycle.

Chemokine Modulation:

According an aspect of some embodiments of the present invention, asmall molecule compound of Formula I, II, III, IV, V, VI, VIA and/orVIB, and/or a compound presented in Table 2, as described herein in anyof the respective embodiments, and any combination thereof, is capableof, or is usable, in modulating a biological activity of a chemokine, asdescribed herein.

According to an aspect of some embodiments of the present invention,there is provided a method of modulating a biological activity of achemokine, the method comprising contacting the chemokine with acompound according to any of the embodiments described herein.

According to an aspect of some embodiments of the present invention,there is provided a use of a compound according to any of theembodiments described herein in the manufacture of a medicament formodulating a biological activity of a chemokine.

According to an aspect of some embodiments of the present invention,there is provided a use of a compound according to any of theembodiments described herein in modulating a biological activity of achemokine.

In some embodiments, the use and/or method for modulating a chemokineactivity is effected in vivo, for example by administering atherapeutically effective amount of the compound to a subject in needthereof.

In some embodiments, the use and/or method for modulating a chemokineactivity is effected ex vivo (e.g., in vitro), for example, in research.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for modulating a biological activity of achemokine, the chemokine is MIP3a, MCP-1 and/or SDF-1.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for modulating a biologicalactivity of a chemokine, the method, use or medicament is for treating adisease or disorder associated with a biological activity of a chemokinein a subject in need thereof, for example, by administering to thesubject a therapeutically effective amount of a compound according toany of the embodiments described herein.

In some of any of the embodiments described herein, modulating abiological activity of a chemokine includes inhibiting a biologicalactivity of a chemokine. This can be evidenced by the ability of a smallmolecule as described herein to inhibit chemokine-induced cell migrationas exemplified herein on a plurality of cell types of different types.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for modulating a biologicalactivity of a chemokine, the method, use or medicament is for treating adisease or disorder in which modulating (e.g., inhibiting) a biologicalactivity of a chemokine is beneficial, in a subject in need thereof, forexample, by administering to the subject a therapeutically effectiveamount of a compound according to any of the embodiments describedherein.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for modulating a biologicalactivity of a chemokine, the method, use or medicament is for treating adisease or disorder treatable by modulating (e.g., inhibiting) abiological activity of a chemokine, in a subject in need thereof, forexample, by administering to the subject a therapeutically effectiveamount of a compound according to any of the embodiments describedherein.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for modulating a biologicalactivity of a chemokine, the compound described herein (according to anyof the respective embodiments) is effective in modulatingchemokine-dependent cell migration. In some of these embodiments, thechemokine-dependent cell migration is associated with cancer and/orinflammation, as described herein.

In some of any of the embodiments described herein, the chemokine isMIP3a.

Examples of diseases and disorders associated with an activity of MIP3a(e.g., wherein inhibition of MIP3a activity is beneficial) include,without limitation, autoimmune diseases and disorders such as psoriasis,inflammatory bowel disease, chronic obstructive pulmonary diseases(COPD), rheumatoid arthritis, multiple sclerosis (MS), atopicdermatitis, dry eye disease and age-related macular degeneration (AMD).

Compounds BKT201, BKT202, BKT203, BKT205, BKT206, BKT207, BKT209,BKT210, BKT212, BKT213 and BKT300 are non-limiting examples of compoundssuitable for inhibiting a MIP3a activity, according to any of therespective embodiments described herein. Compounds BKT202, BKT203,BKT206, BKT207 BKT210, BKT212 and BKT213 are non-limiting examples ofrelatively potent inhibitors of MIP3a activity. Compounds BKT203,BKT206, BKT207 and BKT212 are non-limiting examples of particularlypotent inhibitors of MIP3a activity.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for inhibiting abiological activity of MIP3a, W is an unsubstituted alkyl or alkenyl atleast 5 carbon atoms in length, optionally at least 6 carbon atoms inlength, optionally at least 7 carbon atoms in length, and optionally atleast 8 carbon atoms in length, according to any of the respectiveembodiments described herein.

Compounds BKT205, BKT206, BKT209 and BKT212 are examples of compoundscomprising unsubstituted alkenyl at least 5 carbon atoms in length asthe moiety denoted as W in Formula I. As exemplified herein, CompoundsBKT205, BKT206, BKT209 and BKT212 each exhibited inhibition ofMIP3a-induced cell migration.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for inhibiting abiological activity of MIP3a, W is an unsubstituted alkyl or alkenyl atleast 9 carbon atoms in length, optionally at least 10 carbon atoms inlength, and optionally at least 12 carbon atoms in length, according toany of the respective embodiments described herein.

Compounds BKT206 and BKT212 are examples of compounds comprisingunsubstituted alkenyl at least 9 carbon atoms in length as the moietydenoted as W in Formula I. As exemplified herein, Compounds BKT206 andBKT212 both exhibited potent inhibition of MIP3a-induced cell migration.

In some embodiments of any one of the embodiments described hereinrelating inhibiting a biological activity of MIP3a and/or SDF-1, thecompound has Formula IV or Formula V, according to any of the respectiveembodiments described herein. In some embodiments, R₄-R₅₁ and R₆₁-R₆₈are each hydrogen, hydroxy, alkyl and/or alkylcarboxy.

As exemplified herein, Compound BKT203 (hypericin), which has generalFormula IV, and Compound BKT207, which has general Formula V, areexamples of compounds in which R₄₀-R₅₁ and R₆₁-R₆₈ are each hydrogen,hydroxy, alkyl and/or alkylcarboxy, and are highly effective atinhibiting SDF-1-induced and MIP3a-induced cell migration.

In some embodiments of any one of the embodiments described hereinrelating to a treatment of a disease or disorder, the disease ordisorder is not a bacterial infection.

In some embodiments of any one of the embodiments described hereinrelating to a treatment of a disease or disorder, the disease ordisorder is not a Legionella infection.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for modulating a biological activity of achemokine, the chemokine is MCP-1 and/or SDF-1. In some suchembodiments, the chemokine is MCP-1. In some such embodiments, thechemokine is SDF-1.

In some embodiments of any one of the embodiments described hereinrelating to modulating a chemokine activity, the compound, method and/ormedicament (according to any of the respective embodiments describedherein) is for inhibiting a biological activity of a chemokine. In somesuch embodiments, the chemokine is MCP-1 and/or SDF-1. In some suchembodiments, the chemokine is MCP-1. In some such embodiments, thechemokine is SDF-1.

MCP-1 Inhibition:

According to some embodiments, a small molecule compound of Formula I,II, III, IV, V, VI, VIA and/or VIB, and/or a compound represented inTable 2 herein, as described herein in any of the respectiveembodiments, and any combination thereof, is capable of, or is usable,in modulating a biological activity of MCP-1, as described herein.

According to an aspect of some embodiments of the present invention,there is provided a method of inhibiting a biological activity of MCP-1,the method comprising contacting the MCP-1 with a compound according toany of the embodiments described herein described herein.

According to an aspect of some embodiments of the present invention,there is provided a use of a compound according to any of theembodiments described herein described herein in the manufacture of amedicament for inhibiting a biological activity of MCP-1.

According to an aspect of some embodiments of the present invention,there is provided a use of a compound according to any of theembodiments described herein described herein in inhibiting a biologicalactivity of MCP-1.

In some embodiments of any of the embodiments relating to a use and/ormethod for inhibiting a biological activity of MCP-1, the use and/ormethod is effected in vivo, for example, by administering atherapeutically effective amount of the compound to a subject in needthereof.

In some embodiments, the use and/or method for inhibiting a biologicalactivity of MCP-1 is effected ex vivo (e.g., in vitro), for example, inresearch.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for inhibiting a biologicalactivity of MCP-1, the method, use or medicament is for treating adisease or disorder associated with a biological activity of MCP-1 in asubject in need thereof, for example, by administering to the subject atherapeutically effective amount of a compound according to any of theembodiments described herein described herein.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for inhibiting a biologicalactivity of MCP-1, the method, use or medicament is for treating adisease or disorder in which inhibiting a biological activity of a MCP-1is beneficial, in a subject in need thereof, for example, byadministering to the subject a therapeutically effective amount of acompound according to any of the embodiments described herein describedherein.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for inhibiting a biologicalactivity of MCP-1, the method, use or medicament is for treating adisease or disorder treatable by inhibiting a biological activity of aMCP-1, in a subject in need thereof, for example, by administering tothe subject a therapeutically effective amount of a compound accordingto any of the embodiments described herein described herein.

Examples of diseases and disorders associated with an activity of MCP-1(e.g., wherein inhibition of MCP-1 activity is beneficial) include,without limitation, diseases and disorders which are characterized bymonocytic infiltrates.

According to some embodiments, examples of diseases and disordersassociated with an activity of MCP-1 (e.g., wherein inhibition of MCP-1activity is beneficial) include, without limitation, tuberculosis;HIV-1; proliferative glomerulonephritis; neural tube defects;xanthogranulomatous pyelonephritis; scleritis; rapidly progressiveglomerulonephritis; pneumoconiosis; encephalitis; peritonitis;atherosclerosis; psoriasis; dengue shock syndrome; temporal arteritis;relapsing polychondritis; diabetic angiopathy; mesangial proliferativeglomerulonephritis; sympathetic ophthalmia; ureteral disease; lupusnephritis; pneumonia; periapical granuloma; erdheim-chester disease;glomerulonephritis; artery disease; viral encephalitis; primarycutaneous amyloidosis; arteriosclerosis; nonspecific interstitialpneumonia; acute poststreptococcal glomerulonephritis; coronary arterydisease; venezuelan equine encephalitis; diabetic macular edema;extrapulmonary tuberculosis; nephritis; rheumatoid arthritis; kawasakidisease; arthritis; malaria; obesity; psychiatric disorders; cancer(e.g., as described herein); inflammation (e.g., inflammatory diseaseand disorders as described herein); neurodegenerative disorders; andage-related macular degeneration (AMD, e.g., dry or wet form), asdescribed herein.

According to a specific embodiment, the disease includes, withoutlimitation, psoriasis, rheumatoid arthritis, multiple sclerosis,atherosclerosis, glomerulonephritis, epilepsy, Alzheimer's disease,brain ischemia, traumatic brain injury, type II diabetes and AMD.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for inhibiting abiological activity of MCP-1, Z is —C(═O)OH, Y is absent and L₂ isabsent or is —C(R₂)(OH)—. In some such embodiments, X is cyclic, being,for example, a substituted or unsubstituted bicyclic hydrocarbon moiety,and/or a substituted or unsubstituted phenylene, according to any of therespective embodiments described herein. In some such embodiments, L₂ isabsent.

Compound BKT206 is an example of a compound wherein Z is —C(═O)OH, L₂and Y are each absent, and X is a phenylene. Compounds BKT211 and BKT216are examples of a compound wherein Z is —C(═O)OH, L₂ and Y are eachabsent, and X is a bicyclic hydrocarbon. As exemplified herein,Compounds BKT206, BKT211 and BKT216 each inhibited MCP-1-inducedmigration.

Compounds BKT201, BKT204, BKT205, BKT206, BKT209, BKT211, BKT216 andBKT300 are non-limiting examples of compounds suitable for inhibiting anMCP-1 activity, according to any of the respective embodiments describedherein. Compounds BKT204, BKT206, BKT211 and BKT216 are non-limitingexamples of relatively potent inhibitors of MCP-1 activity, suitable forinhibiting an MCP-1 activity, according to any of the respectiveembodiments described herein. Compound BKT211 is a non-limiting exampleof a particularly potent inhibitor of MCP-1 activity.

Compounds BKT201, BKT204, BKT205, BKT206, BKT211 and BKT300 arenon-limiting examples of compounds suitable for inhibiting MCP-1 inaddition to inhibiting SDF-1, according to any of the respectiveembodiments described herein. Compounds BKT206 and BKT211 arenon-limiting examples of compounds which are relatively potentinhibitors of MCP-1 and SDF-1.

SDF-1 and/or CXCR4 Inhibition:

According to some embodiments, a small molecule compound of Formula I,II, III, IV, V, V, VIA and VIB, and/or a compound presented in Table 2herein, as described herein in any of the respective embodiments, andany combination thereof, is capable of, or is usable, in modulating abiological activity of SDF-1 and/or CXCR4, as described herein.

According to an aspect of some embodiments of the present invention,there is provided a method of inhibiting a biological activity of SDF-1and/or CXCR4, the method comprising contacting the SDF-1 and/or CXCR4with a compound according to any of the embodiments described hereindescribed herein.

According to an aspect of some embodiments of the present invention,there is provided a use of a compound according to any of theembodiments described herein described herein in the manufacture of amedicament for inhibiting a biological activity of SDF-1 and/or CXCR4.

According to an aspect of some embodiments of the present invention,there is provided a use of a compound according to any of theembodiments described herein described herein in inhibiting a biologicalactivity of SDF-1 and/or CXCR4.

In some embodiments of any of the embodiments relating to a use and/ormethod for inhibiting a biological activity of SDF-1 and/or CXCR4, theuse and/or method is effected in vivo, for example, by administering atherapeutically effective amount of the compound to a subject in needthereof.

In some embodiments, the use and/or method for inhibiting a biologicalactivity of SDF-1 and/or CXCR4 is effected ex vivo (e.g., in vitro), forexample, in research.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for inhibiting a biologicalactivity of SDF-1 and/or CXCR4, the method, use or medicament is fortreating a disease or disorder associated with a biological activity ofSDF-1 and/or CXCR4 in a subject in need thereof, for example, byadministering to the subject a therapeutically effective amount of acompound according to any of the embodiments described herein describedherein.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for inhibiting a biologicalactivity of SDF-1 and/or CXCR4, the method, use or medicament is fortreating a disease or disorder in which inhibiting a biological activityof a SDF-1 and/or CXCR4 is beneficial, in a subject in need thereof, forexample, by administering to the subject a therapeutically effectiveamount of a compound according to any of the embodiments describedherein described herein.

In some embodiments of any one of the embodiments described hereinrelating to a method, use or medicament for inhibiting a biologicalactivity of SDF-1 and/or CXCR4, the method, use or medicament is fortreating a disease or disorder treatable by inhibiting a biologicalactivity of a SDF-1 and/or CXCR4 is beneficial, in a subject in needthereof, for example, by administering to the subject a therapeuticallyeffective amount of a compound according to any of the embodimentsdescribed herein described herein.

The skilled person will appreciate that CXCR4 is a receptor whichmediates activity of SDF-1, and that activities of SDF-1 and activitiesof CXCR4 typically overlap.

Examples of diseases and disorders associated with an activity of SDF-1and/or CXCR4 (e.g., wherein inhibition of SDF-1 and/or CXCR4 activity isbeneficial) include, without limitation, Whim Syndrome; CervicalAdenocarcinoma; Breast Cancer; Bursitis; Tuberculosis; IntraocularLymphoma; Cytomegalovirus Retinitis; Chronic Inflammatory DemyelinatingPolyradiculoneuropathy; Ocular Hypertension; Polyradiculoneuropathy;Dendritic Cell Tumor; Retinal Hemangioblastoma; Malaria; Endotheliitis;Leukemia; Rheumatoid Arthritis; Arthritis; Prostatitis; Prostate Cancer;Colorectal Cancer; Chronic Lymphocytic Leukemia; Pancreatitis;Neuronitis; Lung Cancer; Osteoarthritis; Hypoxia; Adenocarcinoma;Pancreatic Cancer; Multiple Myeloma; Neuroblastoma; Myeloid Leukemia;Astrocytoma; Periodontitis; Glioblastoma; Pre-Eclampsia; Melanoma;Hepatitis; Esophagitis; Myeloma; Eclampsia; Cervicitis; PeriodontalDisease; Central Nervous System Lymphoma; Sporadic Breast Cancer;Hepatocellular Carcinoma; Systemic Lupus Erythematosus; Asthma; RenalCell Carcinoma; Myocardial Infarction; Medulloblastoma; EndometrialCancer; Lupus Erythematosus; Esophageal Cancer; Premature OvarianFailure; Peritonitis; Vascular Disease; Alcoholic Hepatitis; KidneyDisease; Cutaneous Leishmaniasis; Encephalitis; Alopecia Areata;Lymphoblastic Leukemia; Adenoma; Mantle Cell Lymphoma;Oligodendroglioma; Malt Lymphoma; Pertussis; Ischemia; Uveal Melanoma;Gingivitis; Pituitary Adenoma; Bronchiolitis; Neuromyelitis Optica;Mesothelioma; Alopecia; Cervical Cancer, Somatic; GlioblastomaMultiforme; Bronchiolitis Obliterans; Brain Injury; Colorectal Adenoma;Tongue Squamous Cell Carcinoma; B-Cell Lymphomas; Traumatic BrainInjury; Intravascular Large B-Cell Lymphoma; Allergic Asthma; Tick-BorneEncephalitis; Blastic Plasmacytoid Dendritic Cell; Oligoastrocytoma;Childhood Type Dermatomyositis; Renal Oncocytoma; EndometrialAdenocarcinoma; Optic Neuritis; Seminoma; Sjogren's Syndrome; Pleurisy;Neuritis; Inflammatory Bowel Disease; Cytomegalovirus Infection;Malignant Pleural Mesothelioma; Oral Squamous Cell Carcinoma; SkeletalMuscle Regeneration; Emery-Dreifuss Muscular Dystrophy, Dominant Type.

In some embodiments, exemplary diseases and disorders associated with anactivity of SDF-1 and/or CXCR4 (e.g., wherein inhibition of SDF-1 and/orCXCR4 activity is beneficial) include, without limitation, harmfulangiogenesis, tumor metastasis, WHIM syndrome, Waldenstrommacroglobulinemia (WM) and opioid-induced hyperalgesia.

Herein, the term “harmful angiogenesis” refers to angiogenesisassociated with a clinically and/or cosmetically undesirable result.

Angiogenesis associated with a tumor is a non-limiting example of aharmful angiogenesis.

As used herein the phrase “tumor metastasis” refers to a malignant tumorspreading out of its primary location to other parts of the body, e.g.,breast cancer which metastasizes to the lungs. Tumor metastasis ofteninvolves migration of tumor cells.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for modulating a biological activity of achemokine, the modulating comprises inhibiting a biological activity ofSDF-1 and/or CXCR4, according to any of the respective embodimentsdescribed herein.

In some embodiments of any one of the embodiments described hereinrelating to inhibiting a biological activity of SDF-1 and/or CXCR4,inhibiting a biological activity of SDF-1 and/or CXCR4 is for effectingimmunostimulation.

In some embodiments, immunostimulation is effected as part of a cancertreatment, e.g., in order to stimulate immune activity against cancercells.

In some embodiments, immunostimulation comprises increasing a level ofhematopoietic stem cells in peripheral blood of a subject.

In some embodiments, increasing a level of hematopoietic stem cells inperipheral blood of a subject is effected as a preliminary part ofhematopoietic stem cell transplantation (e.g., in order to generatehematopoietic stem cells available for collection and latertransplantation back into the subject). Examples of conditions which maybe treated by the hematopoietic stem cell transplantation include,without limitation, leukemia (e.g., acute lymphoblastic leukemia, acutemyeloid leukemia, chronic lymphocytic leukemia, chronic myelogenousleukemia), lymphoma (e.g., Hodgkin's disease, non-Hodgkin's lymphoma),myeloma (e.g., multiple myeloma), neuroblastoma, desmoplastic smallround cell tumor, Ewing's sarcoma, choriocarcinoma, myelodysplasia,anemias (e.g., paroxysmal nocturnal hemoglobinuria, aplastic anemia,Diamond-Blackfan anemia, Fanconi anemia, acquired pure red cellaplasia), hemoglobinopathies, sickle cell disease, beta-thalassemiamajor, myeloproliferative disorders (e.g., polycythemia vera, essentialthrombocytosis, myelofibrosis), amyloid light chain amyloidosis,radiation poisoning, viral diseases (e.g., HTLV and/or HIC infection),neuronal ceroid lipofuscinosis, Niemann-Pick disease, Gaucher disease,leukodystrophies (adrenoleukodystrophy, metachromatic leukodystrophy,Krabbe disease), mucopolysaccharoidosis, glycoproteinoses (e.g.,mucolipidosis II, fucosidosis, aspartylglucosaminuria,alpha-mannosidosis), Wolman disease, immunodeficiencies (e.g., ataxiatelangiectasia, DiGeorge syndrome, severe combined immunodeficiency,Wiskott-Aldrich syndrome, Kostmann syndrome, Shwachman-Diamond syndrome,Griscelli syndrome, NF-kappa-B essential modulator deficiency),amegakaryocytic thrombocytopenia and hemophagocytic lymphohistiocytosis.

In some embodiments, the hematopoietic stem cell transplantation is fortreating a proliferative disease, e.g., cancer (e.g., cancer asdescribed herein according to any of the respective embodiments).

In some embodiments of any one of the embodiments described hereinrelating to hematopoietic stem cells, the treatment comprises increasinga level of hematopoietic stem cells in peripheral blood of the subject,obtaining hematopoietic stem cells from peripheral blood of the subject,administering a cytotoxic therapy to the subject (e.g.,anti-proliferative chemotherapy, and/or radiotherapy), and transplantingat least a portion of the stem cells back into the patient, subsequentto the cytotoxic therapy.

Compounds presented in Table 2 are non-limiting examples of compoundswhich may be used for inhibiting a biological activity of SDF-1 and/orCXCR4 according to any of the respective embodiments described herein.In some such embodiments, the compound is Compound BKT204 in Table 2.

Compound BKT201, BKT203, BKT204, BKT205, BKT206, BKT207, BKT208, BKT209,BKT210, BKT211, BKT212, BKT213, BKT300 and BKT400 are non-limitingexamples of compounds suitable for inhibiting an SDF-1 activity,according to any of the respective embodiments described herein.Compounds BKT201, BKT203, BKT205, BKT206, BKT207, BKT209, BKT211,BKT212, BKT213 and BKT300 are non-limiting examples of compounds whichexhibit particularly potent inhibition of SDF-1 activity. CompoundsBKT201, BKT205, BKT209, BKT213 and BKT300 are non-limiting examples ofcompounds which exhibit particularly potent and relatively selectiveinhibition of SDF-1 activity.

Compounds BKT201, BKT203, BKT205, BKT206, BKT207, BKT209, BKT210,BKT212, BKT213 and BKT300 are non-limiting examples of compoundssuitable for inhibiting MIP3a in addition to inhibiting SDF-1, accordingto any of the respective embodiments described herein. Compounds BKT203,BKT206, BKT207 or BKT212 are non-limiting examples of potent inhibitorsof both MIP3a and SDF-1.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for inhibiting abiological activity of SDF-1 and/or CXCR4, L₂ in Formula I is absent, oralternatively, is —C(═O)— or —C(R₂)(OH)—, in accordance with any of therespective embodiments described herein.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for inhibiting abiological activity of SDF-1 and/or CXCR4, Z is —C(═O)OH.

In some embodiments, Z is —C(═O)OH, Y is absent and L₂ is absent or is—C(R₂)(OH)—. In some such embodiments, X is cyclic, being, for example,a substituted or unsubstituted bicyclic hydrocarbon moiety, and/or asubstituted or unsubstituted phenylene, according to any of therespective embodiments described herein. In some such embodiments, L₂ isabsent.

Compound BKT206 is an example of a compound wherein Z is —C(═O)OH, L₂and Y are each absent, and X is a phenylene. Compound BKT211 is anexample of a compound wherein Z is —C(═O)OH, L₂ and Y are each absent,and X is a bicyclic hydrocarbon. As exemplified herein, Compounds BKT206and BKT211 each exhibited SDF-1-induced cell migration.

Compound BKT213 is an example of a compound wherein Z is —C(═O)OH, Y isabsent, L₂ is —C(R₂)(OH)—, and X is a phenylene. As exemplified herein,Compound BKT213 inhibited SDF-1-induced cell migration.

In some embodiments, Z is —C(═O)OH, Y is an aliphatic hydrocarbonmoiety, and X is —C≡C—C≡C— or —CR₃═CR₄—CR₅═CR₆—, according to any of therespective embodiments described herein. In some embodiments, X is—CR₃═CR₄—CR₅═CR₆—. In some embodiments, L₂ is —C(═O)—. In someembodiments, L₂ is —C(═O)— and X is —CR₃═CR₄—CR₅═CR₆—.

Compounds BKT205 and BKT209 are examples of compound wherein Z is—C(═O)OH, Y is an aliphatic hydrocarbon moiety (a linear andunsubstituted saturated hydrocarbon moiety) from 1 to 8 atoms in length,L₂ is —C(═O)— and X is —CR₃═CR₄—CR₅═CR₆—. As exemplified herein,Compounds BKT205 and BKT209 each inhibited SDF-1-induced cell migration.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for inhibiting abiological activity of SDF-1 and/or CXCR4, X is —C≡C—C≡C— or—CR₃═CR₄—CR₅═CR₆—, according to any of the respective embodimentsdescribed herein. In some embodiments, Y is an aliphatic hydrocarbonmoiety.

In some embodiments, X is —C≡C—C≡C—, according to any of the respectiveembodiments described herein. In some embodiments, Z is absent and/or L₁and L₂ are each —CH(OH)—.

Compound BKT212 (falcarindiol) is an example of a compound in which X is—C≡C—C≡C—, Z is absent and L₁ and L₂ are each —CH(OH)—. As exemplifiedherein, Compound BKT212 inhibited SDF-1-induced cell migration.

In some embodiments, X is —CR₃═CR₄—CR₅═CR₆—, according to any of therespective embodiments described herein. In some embodiments, L₂ is—C(═O)—.

Compounds BKT205 and BKT209 are examples of compounds wherein X is—CR₃═CR₄—CR₅═CR₆—, and as exemplified herein, Compounds BKT205 andBKT209 each inhibited SDF-1-induced cell migration.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula II for inhibiting abiological activity of SDF-1 and/or CXCR4, Q is —C(═O)—. In some suchembodiments, J is —O— or —CH═, optionally —O—. In some such embodiments,K is —CR₁₆R₁₇— wherein R₁₆ is alkyl or phenyl, each being substituted orunsubstituted.

Compounds BKT201, BKT204 and BKT300 are examples of compounds in which Qis —C(═O)—. As exemplified herein, each of compounds BKT201, BKT204 andBKT300 inhibited SDF-1-induced cell migration to a considerable extent.

In some embodiments of any one of the embodiments described hereinrelating inhibiting a biological activity of SDF-1 and/or CXCR4, thecompound has Formula IV or Formula V, according to any of the respectiveembodiments described herein. In some embodiments, R₄-R₅₁ and R₆₁-R₆₈are each hydrogen, hydroxy, alkyl and/or alkylcarboxy.

Kinase Inhibition:

According to some of any of the embodiments described herein, a compoundrepresented by Formula I, II, III, IV, V, VI, VIA and/or VIB hereinand/or a compound presented in Table 2, is capable of, or is usable in,inhibiting a biological activity of a kinase. In some such embodiments,the compound is represented Formula VIA and/or VIB herein.

According to some of any of the embodiments described herein, a compoundrepresented Formula I, II, III, IV, V, VI, VIA and/or VIB herein and/ora compound presented in Table 2, is capable of, or is usable in,treating diseases or disorder in which inhibiting a biological activityof a kinase is beneficial, or a disease or disorder that is treatable byinhibiting a biological activity of a kinase. In some such embodiments,the compound is represented Formula VIA and/or VIB herein.

According to an aspect of some embodiments of the present invention, acompound according to any of the embodiments described herein, is foruse in inhibiting a biological activity of a kinase. In some suchembodiments, the compound is represented Formula VIA and/or VIB herein.

According to another aspect of some embodiments of the presentinvention, there is provided a method of inhibiting a biologicalactivity of a kinase, the method comprising contacting the kinase with acompound according to any of the embodiments described herein. In somesuch embodiments, the compound is represented Formula VIA and/or VIBherein.

In some embodiments, the use and/or method for inhibiting a kinase iseffected ex vivo (e.g., in vitro), for example, in research.

In some embodiments, the use and/or method for inhibiting a kinase iseffected in vivo, for example by administering a therapeuticallyeffective amount of the compound to a subject in need thereof.

According to another aspect of some embodiments of the presentinvention, there is provided a use of a compound according to any of theembodiments described herein in the manufacture of a medicament for usein inhibiting a biological activity of a kinase in a subject in needthereof. In some such embodiments, the compound is represented FormulaVIA and/or VIB herein.

In some embodiments of any one of the embodiments described hereinrelating to a use, method and/or medicament for inhibiting a biologicalactivity of a kinase, the use, method and/or medicament (according toany of the respective embodiments described herein) is for use intreating a disease or disorder associated with a biological activity ofa kinase in a subject in need thereof.

In some embodiments of any one of the embodiments described hereinrelating to a use, method and/or medicament for inhibiting a biologicalactivity of a kinase, the use, method and/or medicament is for use intreating a disease or disorder in which inhibition of a biologicalactivity of a kinase is beneficial.

In some embodiments of any one of the embodiments described hereinrelating to a use, method and/or medicament for inhibiting a biologicalactivity of a kinase, the use, method and/or medicament is for treatinga disease or disorder which is treatable by inhibition of a biologicalactivity of a kinase.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for inhibiting a biological activity of akinase, the inhibited kinase can be a kinase presented in Table 3 below,for example, DYRK3, EPHA8, GRK4, GRK5, MAP4K1, MAP4K2, MAP4K4, MELK,PAK7, SGK2, SRC N1, ACVRL1, BMPR1A, CDCl₇/DBF4, CDK1/cyclin A2, CDK11,CDK8/cyclin C, CLK4, DAPK2, DURK2, ICK, MAPK10, MLCK, MYLK, NUAK2,STK17A, STK17B, STK38, STK38L, TGFBR2, TTK, DAPK1, PIK3CA and/or PIK3CD.

According to a specific embodiment, the kinase is a PI3K.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for inhibiting a biological activity of akinase, the inhibited kinase is a serine/threonine kinase. In someembodiments, the serine/threonine kinase is a serine/threonine kinasepresented in Table 3 below.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for inhibiting a biological activity of akinase, the inhibited kinase is a tyrosine kinase. In some embodiments,the tyrosine kinase is a serine/threonine kinase presented in Table 3below.

In some embodiments of any one of the embodiments described hereinrelating to a method or use for inhibiting a biological activity of akinase, the kinase is MELK, MAP4K4 and/or PI3K.

Cancer Treatment:

According to some embodiments, a small molecule compound of Formula I,II, III, IV, V, VI, VIA and/or VIB, and/or a compound presented in Table2, as described herein in any of the respective embodiments, and anycombination thereof, is capable of, or is usable, in treating cancer.

According to some embodiments, a small molecule compound of Formula I,II, III, IV, V, VI, VIA and/or VIB, and/or a compound presented in Table2, as described herein in any of the respective embodiments, and anycombination thereof, is capable of, or is usable, in inducing death ofcancer cells (killing cancer cells).

According to some embodiments, a small molecule compound of Formula I,II, III, IV, V, VI, VIA and/or VIB, and/or a compound presented in Table2, as described herein in any of the respective embodiments, and anycombination thereof, is capable of, or is usable, inducing apoptosis incancer cells.

According to some embodiments, a small molecule compound of Formula I,II, III, IV, V, VI, VIA and/or VIB, and/or a compound presented in Table2, as described herein in any of the respective embodiments, and anycombination thereof, is capable of, or is usable, in inducing growtharrest in cancer cells, and in some embodiments, the arrest is at theG2M phase of the cell cycle.

According to an aspect of some embodiments of the present invention,there is provided a method of treating a cancer in a subject in needthereof, the method comprising administering to the subject atherapeutically effective amount of a small molecule compound accordingto any of the embodiments described herein, thereby treating the cancer.

According to an aspect of some embodiments of the present invention,there is provided a use of a small molecule compound according to any ofthe embodiments described herein in the manufacture of a medicament fortreating cancer.

According to an aspect of some embodiments of the present invention,there is provided a use of a small molecule compound according to any ofthe embodiments described herein in treating cancer.

As used herein, the terms “cancer” and “tumor” are interchangeably used.The terms refer to a malignant growth and/or tumor caused by abnormaland uncontrolled cell proliferation (cell division). The term “cancer”encompasses tumor metastases. The term “cancer cell” describes the cellsforming the malignant growth or tumor.

Non-limiting examples of cancers and/or tumor metastases which can betreated according to some embodiments of any of the embodimentsdescribed herein relating to cancer (including any of the aspectsdescribed herein) include any solid or non-solid cancer and/or tumormetastasis, including, but not limiting to, tumors of thegastrointestinal tract (e.g., colon carcinoma, rectal carcinoma,colorectal carcinoma, colorectal cancer, colorectal adenoma, hereditarynonpolyposis type 1, hereditary nonpolyposis type 2, hereditarynonpolyposis type 3, hereditary nonpolyposis type 6; colorectal cancer,hereditary nonpolyposis type 7, small and/or large bowel carcinoma,esophageal carcinoma, tylosis with esophageal cancer, stomach carcinoma,pancreatic carcinoma, pancreatic endocrine tumors), endometrialcarcinoma, dermatofibrosarcoma protuberans, gallbladder carcinoma,biliary tract tumors, prostate cancer, prostate adenocarcinoma, renalcancer (e.g., Wilms' tumor type 2 or type 1), liver cancer (e.g.,hepatoblastoma, hepatocellular carcinoma, hepatocellular cancer),bladder cancer, embryonal rhabdomyosarcoma, germ cell tumor,trophoblastic tumor, testicular germ cells tumor, immature teratoma ofovary, uterine, epithelial ovarian, sacrococcygeal tumor,choriocarcinoma, placental site trophoblastic tumor, epithelial adulttumor, ovarian carcinoma, serous ovarian cancer, ovarian sex cordtumors, cervical carcinoma, uterine cervix carcinoma, small-cell andnon-small cell lung carcinoma, nasopharyngeal, breast carcinoma (e.g.,ductal breast cancer, invasive intraductal breast cancer, sporadicbreast cancer, susceptibility to breast cancer, type 4 breast cancer,breast cancer-1, breast cancer-3, breast-ovarian cancer), squamous cellcarcinoma (e.g., in head and neck), neurogenic tumor, astrocytoma,ganglioblastoma, neuroblastoma, lymphomas (e.g., Hodgkin's disease,non-Hodgkin's lymphoma, B-cell lymphoma, Diffuse large B-cell lymphoma(DLBCL), Burkitt lymphoma, cutaneous T-cell lymphoma, histiocyticlymphoma, lymphoblastic lymphoma, T-cell lymphoma, thymic lymphoma),gliomas, adenocarcinoma, adrenal tumor, hereditary adrenocorticalcarcinoma, brain malignancy (tumor), various other carcinomas (e.g.,bronchogenic large cell, ductal, Ehrlich-Lettre ascites, epidermoid,large cell, Lewis lung, medullary, mucoepidermoid, oat cell, small cell,spindle cell, spinocellular, transitional cell, undifferentiated,carcinosarcoma, choriocarcinoma, cystadenocarcinoma), ependimoblastoma,epithelioma, erythroleukemia (e.g., Friend, lymphoblast), fibrosarcoma,giant cell tumor, glial tumor, glioblastoma (e.g., multiforme,astrocytoma), glioma hepatoma, heterohybridoma, heteromyeloma,histiocytoma, hybridoma (e.g., B-cell), hypernephroma, insulinoma, islettumor, keratoma, leiomyoblastoma, leiomyosarcoma, leukemia (e.g., acutelymphatic leukemia, acute lymphoblastic leukemia, acute lymphoblasticpre-B cell leukemia, acute lymphoblastic T cell leukemia, acutemegakaryoblastic leukemia, monocytic leukemia, acute myelogenousleukemia, acute myeloid leukemia, acute myeloid leukemia witheosinophilia, B-cell leukemia, basophilic leukemia, chronic myeloidleukemia, chronic B-cell leukemia, eosinophilic leukemia, Friendleukemia, granulocytic or myelocytic leukemia, hairy cell leukemia,lymphocytic leukemia, megakaryoblastic leukemia, monocytic leukemia,monocytic-macrophage leukemia, myeloblastic leukemia, myeloid leukemia,myelomonocytic leukemia, plasma cell leukemia, pre-B cell leukemia,promyelocytic leukemia, subacute leukemia, T-cell leukemia, lymphoidneoplasm, predisposition to myeloid malignancy, acute nonlymphocyticleukemia), lymphosarcoma, melanoma, mammary tumor, mastocytoma,medulloblastoma, mesothelioma, metastatic tumor, monocyte tumor,multiple myeloma, myelodysplastic syndrome, myeloma, nephroblastoma,nervous tissue glial tumor, nervous tissue neuronal tumor, neurinoma,neuroblastoma, oligodendroglioma, osteochondroma, osteomyeloma,osteosarcoma (e.g., Ewing's), papilloma, transitional cell,pheochromocytoma, pituitary tumor (invasive), plasmacytoma,retinoblastoma, rhabdomyosarcoma, sarcoma (e.g., Ewing's, histiocyticcell, Jensen, osteogenic, reticulum cell), schwannoma, subcutaneoustumor, teratocarcinoma (e.g., pluripotent), teratoma, testicular tumor,thymoma and trichoepithelioma, gastric cancer, fibrosarcoma,glioblastoma multiforme, multiple glomus tumors, Li-Fraumeni syndrome,liposarcoma, lynch cancer family syndrome II, male germ cell tumor, mastcell leukemia, medullary thyroid, multiple meningioma, endocrineneoplasia myxosarcoma, paraganglioma, familial nonchromaffin,pilomatricoma, papillary, familial and sporadic, rhabdoid predispositionsyndrome, familial, rhabdoid tumors, soft tissue sarcoma, and Turcotsyndrome with glioblastoma.

In some embodiments of any one of the embodiments described hereinrelating to cancer, the cancer is a leukemia, a lymphoma, ovariancancer, neuroblastoma, a prostate cancer and/or a lung cancer.

Examples of leukemias which may be treated in the context of someembodiments of the invention include, without limitation, acuteleukemias, for example, acute myeloid leukemia (AML), chronic myeloidleukemia (CML) and acute lymphoblastic leukemia.

Examples of lymphomas which may be treated in the context of someembodiments of the invention include, without limitation, diffuse largeB-cell lymphoma (DLBCL), multiple myeloma and non-Hodgkin's lymphomas.Burkitt lymphoma is a non-limiting example of a non-Hodgkin's lymphoma.

Examples of lung cancers which may be treated in the context of someembodiments of the invention include, without limitation, large celllung cancer and small cell lung cancer.

In some embodiments of any one of the embodiments described hereinrelating to cancer, the cancer is characterized by cells expressingCXCR4. In some such embodiments, the compound for use in treating canceris any one of the compounds described herein as being for use ininhibiting SDF-1 and/or CXCR4 activity.

Without being bound by any particular theory, it is believed that incancers characterized by expression of CXCR4, the activity of SDF-1 andCXCR4 is generally associated with metastasis, and thus, treatment withan inhibitor of SDF-1 and/or CXCR4 activity is particularlyadvantageous.

In some embodiments of any one of the embodiments described hereinrelating to treatment of cancer, the cancer further comprisesadministering at least one additional anti-cancer agent (i.e., inaddition to the compound described hereinabove).

The additional anti-cancer agent may be any agent used in the medicalarts to treat a cancer. Examples of anti-cancer agents include, withoutlimitation, acivicin; aclarubicin; acodazole hydrochloride; acronine;adriamycin; Adozelesin; aldesleukin; altretamine; ambomycin; ametantroneacetate; aminoglutethimide; amsacrine; anastrozole; anthramycin;asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat;benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate;bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cisplatin; cladribine; combrestatin A-4phosphate; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine;dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel;doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifenecitrate; dromostanolone propionate; duazomycin; edatrexate; eflornithinehydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine;epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;estramustine; estramustine phosphate sodium; etanidazole; etoposide;etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine;fenretinide; floxuridine; fludarabine phosphate; fluorouracil;flurocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabinehydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;ilmofosine; interferon alfa-2a; interferon alfa-2b; interferon alfa-n1;interferon alfa-n3; interferon beta-Ia; interferon gamma-Ib; iproplatin;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; ombrabulin; ormaplatin; oxisuran;paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantronehydrochloride; temoporfin; teniposide; teroxirone; testolactone;thiamiprine; thioguanine; thiotepa; tiazofuirin; tirapazamine; topotecanhydrochloride; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine; vincristine sulfate; vindesine; vindesinesulfate; vinepidinee; vinglycinate; vinleurosine; vinorelbine tartrate;vinrosidine; vinzolidine; vorozole; zeniplatin; zinostatin; andzorubicin hydrochloride. Additional anti-cancer agents include thosedisclosed in Chapter 52, Antineoplastic Agents (Paul Calabresi and BruceA. Chabner), and the introduction thereto, 1202-1263, of Goodman andGilman's “The Pharmacological Basis of Therapeutics”, Eighth Edition,1990, McGraw-Hill, Inc. (Health Professions Division), the contents ofwhich are incorporated herein by reference.

In some embodiments of any of the embodiments described herein, theadditional anti-cancer agent is characterized in that resistance ofcancer cells to the agent is associated with an activity of SDF-1 and/orCXCR4 and/or any one of the kinases described in Table 1 herein. In somesuch embodiments, the compound for use in combination with theadditional anti-cancer agent is any one of the compounds describedherein.

In some embodiments of any of the embodiments described herein, the atleast one additional anti-cancer agent comprises combrestatin A-4phosphate, ombrabulin and/or any other derivative of combrestatin.

Without being bound by any particular theory, it is believed that theanti-therapeutic effect of combrestatin derivatives such as combrestatinA-4 phosphate and ombrabulin is reduced by SDF-1/CXCR4 activity.

In some embodiments of any of the embodiments described herein relatingto treating a cancer, treating a cancer does not comprise illuminatingthe compound in situ (within or on a surface of the body of the subject)at a wavelength absorbed by the compound, e.g., as in photodynamictherapy.

Herein, the term “illuminating” refers to irradiating a target (e.g., abody of a subject) with electromagnetic radiation having a wavelength ina range of from 300 to 1000 nm.

The compounds presented in Table 2 are non-limiting examples ofcompounds which may be used for treating cancer according to any of therespective embodiments described herein. In some such embodiments, thecompound is Compound BKT206, BKT211, BKT215, or BKT300, as presented inTable 2. In some embodiments, the compound is Compound BKT206, BKT211,or BKT300. In some embodiments, the compound is Compound BKT204, BKT214,or BKT300. In some embodiments, the compound is Compound BKT204 orBKT214.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for treating acancer, L₂ in Formula I is absent or is —C(═O)— or —C(R₂)(OH)—, inaccordance with any of the respective embodiments described herein.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for treating acancer, X in Formula I is —CR₃═CR₄—CR₅═CR₆—, a substituted orunsubstituted bicyclic hydrocarbon moiety or substituted orunsubstituted phenylene, in accordance with any of the respectiveembodiments described herein.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for treating acancer, L₂ in Formula I is absent or is —C(═O)— or —C(R₂)(OH)—, and X inFormula I is —CR₃═CR₄—CR₅═CR₆—, a substituted or unsubstituted bicyclichydrocarbon moiety, or substituted or unsubstituted phenylene, inaccordance with any of the respective embodiments described herein.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula I for treating acancer, Z is —C(═O)OH, Y is absent and L₂ is absent or is —C(R₂)(OH)—.In some such embodiments, X is cyclic, being, for example, a substitutedor unsubstituted bicyclic hydrocarbon moiety, and/or a substituted orunsubstituted phenylene, according to any of the respective embodimentsdescribed herein. In some such embodiments, L₂ is absent.

Compound BKT206 is an example of a compound wherein Z is —C(═O)OH, L₂and Y are each absent, and X is a phenylene. Compound BKT211 is anexample of a compound wherein Z is —C(═O)OH, L₂ and Y are each absent,and X is a bicyclic hydrocarbon. As exemplified herein, Compounds BKT206and BKT211 each exhibited anticancer activity in a leukemia cell model.

In some embodiments of any one of the embodiments described hereinrelating to use of a compound having general Formula II for treatingcancer, Q is —C(═O)—. In some such embodiments, J is —O— or —CH═,optionally —O—. In some such embodiments, K is —CR₁₆R₁₇— wherein R₁₆ isalkyl or phenyl, each being substituted or unsubstituted. CompoundsBKT201, BKT204 and BKT300 (e.g., 78% and 98% purity) are examples ofcompounds in which Q is —C(═O)—. As exemplified herein, Compound BKT300exhibited anticancer activity in a leukemia cell model.

Non-Cancerous Hyperproliferative Diseases:

Non-cancerous hyperproliferative diseases also referred to“non-neoplastic diseases” refer to diseases or disorders which onset orprogression is associated with non-malignant cell proliferation.Examples of such medical conditions include, but are not limited toatherosclerosis, rheumatoid arthritis, psoriasis, fibrosis, idiopathicpulmonary fibrosis, scleroderma and cirrhosis of the live.

Inflammatory Diseases and Disorders:

Inflammatory diseases and disorders generally encompass diseases anddisorders associated with inflammation.

The term “inflammation” as used herein refers to the general term forlocal accumulation of fluids, plasma proteins, and white blood cellsinitiated by physical injury, infection, or a local immune response.Inflammation may be associated with several signs e.g. redness, pain,heat, swelling and/or loss of function. Inflammation is an aspect ofmany diseases and disorders, including but not limited to diseasesrelated to immune disorders, viral and bacterial infection, arthritis,autoimmune diseases, collagen diseases, allergy, asthma, pollinosis, andatopy (as described in further detail below).

Thus, inflammation can be triggered by injury, for example injury toskin, muscle, tendons, or nerves. Inflammation can be triggered as partof an immune response, e.g., pathologic autoimmune response.Inflammation can also be triggered by infection, where pathogenrecognition and tissue damage can initiate an inflammatory response atthe site of infection.

Inflammation according to the present teachings may be associated withchronic (long term) inflammatory diseases or disorders or acute (shortterm) inflammatory diseases or disorders.

According to a specific embodiment, the inflammation is associated witha disease selected from the group consisting of an infectious disease,an autoimmune disease, a hypersensitivity associated inflammation, agraft rejection and an injury.

According to a specific embodiment, the inflammation comprises a skininflammation.

According to a specific embodiment the skin inflammation is psoriasis.

Diseases characterized by inflammation of the skin, include but are notlimited to dermatitis, atopic dermatitis (eczema, atopy), contactdermatitis, dermatitis herpetiformis, generalized exfoliativedermatitis, seborrheic dermatitis, drug rashes, erythema multiforme,erythema nodosum, granuloma annulare, poison ivy, poison oak, toxicepidermal necrolysis, roseacae, psoriasis and acne. Inflammation canalso result from physical injury to the skin.

Inflammation may be triggered by various kinds of injuries to muscles,tendons or nerves. Thus, for example, inflammation may be caused byrepetitive movement of a part of the body i.e. repetitive strain injury(RSI). Diseases characterized by inflammation triggered by RSI include,but are not limited to, bursitis, carpal tunnel syndrome, Dupuytren'scontracture, epicondylitis (e.g. tennis elbow), ganglion (i.e.inflammation in a cyst that has formed in a tendon sheath, usuallyoccurring on the wrist), rotator cuff syndrome, tendinitis (e.g.,inflammation of the Achilles tendon), tenosynovitis, and trigger finger(inflammation of the tendon sheaths of fingers or thumb accompanied bytendon swelling).

Many diseases related to infectious diseases include inflammatoryresponses, where the inflammatory responses are typically part of theinnate immune system triggered by the invading pathogen. Inflammationcan also be triggered by physical (mechanical) injury to cells andtissues resulting from the infection. Examples of infectious diseasesinclude, but are not limited to, chronic infectious diseases, subacuteinfectious diseases, acute infectious diseases, viral diseases,bacterial diseases, protozoan diseases, parasitic diseases, fungaldiseases, mycoplasma diseases and prion diseases. According to oneembodiment, examples of infections characterized by inflammationinclude, but are not limited to, encephalitis; meningitis;encephalomyelitis; viral gastroenteritis; viral hepatitis.

Furthermore, many immune disorders include acute or chronicinflammation. For example, arthritis is considered an immune disordercharacterized by inflammation of joints, but arthritis is likewiseconsidered an inflammatory disorder characterized by immune attack onjoint tissues.

Inflammation according to the present teachings may be associated with adeficient immune response (e.g., HIV, AIDS) or with an overactive immuneresponse (e.g., allergy, autoimmune disorders). Thus, inflammationaccording to the present teachings may be associated with any of thefollowing:

Inflammatory Diseases Associated with Hypersensitivity:

Examples of hypersensitivity include, but are not limited to, Type Ihypersensitivity, Type II hypersensitivity, Type III hypersensitivity,Type IV hypersensitivity, immediate hypersensitivity, antibody mediatedhypersensitivity, immune complex mediated hypersensitivity, T lymphocytemediated hypersensitivity and DTH.

Type I or immediate hypersensitivity, such as asthma.

Type II hypersensitivity include, but are not limited to, rheumatoiddiseases, rheumatoid autoimmune diseases, rheumatoid arthritis (Krenn V.et al., Histol Histopathol 2000 July; 15 (3):791), spondylitis,ankylosing spondylitis (Jan Voswinkel et al., Arthritis Res 2001; 3 (3):189), systemic diseases, systemic autoimmune diseases, systemic lupuserythematosus (Erikson J. et al., Immunol Res 1998; 17 (1-2):49),sclerosis, systemic sclerosis (Renaudineau Y. et al., Clin Diagn LabImmunol. 1999 March; 6 (2):156); Chan OT. et al., Immunol Rev 1999 June;169:107), glandular diseases, glandular autoimmune diseases, pancreaticautoimmune diseases, diabetes, Type I diabetes (Zimmet P. Diabetes ResClin Pract 1996 October; 34 Suppl:S125), thyroid diseases, autoimmunethyroid diseases, Graves' disease (Orgiazzi J. Endocrinol Metab ClinNorth Am 2000 June; 29 (2):339), thyroiditis, spontaneous autoimmunethyroiditis (Braley-Mullen H. and Yu S, J Immunol 2000 Dec. 15; 165(12):7262), Hashimoto's thyroiditis (Toyoda N. et al., Nippon Rinsho1999 August; 57 (8):1810), myxedema, idiopathic myxedema (Mitsuma T.Nippon Rinsho. 1999 August; 57 (8):1759); autoimmune reproductivediseases, ovarian diseases, ovarian autoimmunity (Garza KM. et al., JReprod Immunol 1998 February; 37 (2):87), autoimmune anti-sperminfertility (Diekman AB. et al., Am J Reprod Immunol. 2000 March; 43(3):134), repeated fetal loss (Tincani A. et al., Lupus 1998; 7 Suppl2:S107-9), neurodegenerative diseases, neurological diseases,neurological autoimmune diseases, multiple sclerosis (Cross A H. et al.,J Neuroimmunol 2001 Jan. 1; 112 (1-2):1), Alzheimer's disease (Oron L.et al., J Neural Transm Suppl. 1997; 49:77), myasthenia gravis (InfanteA J. And Kraig E, Int Rev Immunol 1999; 18 (1-2):83), motor neuropathies(Kornberg A J. J Clin Neurosci. 2000 May; 7 (3):191), Guillain-Barresyndrome, neuropathies and autoimmune neuropathies (Kusunoki S. Am J MedSci. 2000 April; 319 (4):234), myasthenic diseases, Lambert-Eatonmyasthenic syndrome (Takamori M. Am J Med Sci. 2000 April; 319 (4):204),paraneoplastic neurological diseases, cerebellar atrophy, paraneoplasticcerebellar atrophy, non-paraneoplastic stiff man syndrome, cerebellaratrophies, progressive cerebellar atrophies, encephalitis, Rasmussen'sencephalitis, amyotrophic lateral sclerosis, Sydeham chorea, Gilles dela Tourette syndrome, polyendocrinopathies, autoimmunepolyendocrinopathies (Antoine J C. and Honnorat J. Rev Neurol (Paris)2000 January; 156 (1):23); neuropathies, dysimmune neuropathies(Nobile-Orazio E. et al., Electroencephalogr Clin Neurophysiol Suppl1999; 50:419); neuromyotonia, acquired neuromyotonia, arthrogryposismultiplex congenita (Vincent A. et al., Ann N Y Acad Sci. 1998 May 13;841:482), cardiovascular diseases, cardiovascular autoimmune diseases,atherosclerosis (Matsuura E. et al., Lupus. 1998; 7 Suppl 2:S135),myocardial infarction (Vaarala O. Lupus. 1998; 7 Suppl 2:S132),thrombosis (Tincani A. et al., Lupus 1998; 7 Suppl 2:S107-9),granulomatosis, Wegener's granulomatosis, arteritis, Takayasu'sarteritis and Kawasaki syndrome (Praprotnik S. et al., Wien KlinWochenschr 2000 Aug. 25; 112 (15-16):660); anti-factor VIII autoimmunedisease (Lacroix-Desmazes S. et al., Semin Thromb Hemost.2000; 26(2):157); vasculitises, necrotizing small vessel vasculitises,microscopic polyangiitis, Churg and Strauss syndrome,glomerulonephritis, pauci-immune focal necrotizing glomerulonephritis,crescentic glomerulonephritis (Noel L H. Ann Med Internet (Paris). 2000May; 151 (3):178); antiphospholipid syndrome (Flamholz R. et al., J ClinApheresis 1999; 14 (4):171); heart failure, agonist-like β-adrenoceptorantibodies in heart failure (Wallukat G. et al., Am J Cardiol. 1999 Jun.17; 83 (12A):75H), thrombocytopenic purpura (Moccia F. Ann Ital Med Int.1999 April-June; 14 (2):114); hemolytic anemia, autoimmune hemolyticanemia (Efremov DG. et al., Leuk Lymphoma 1998 January; 28 (3-4):285),gastrointestinal diseases, autoimmune diseases of the gastrointestinaltract, intestinal diseases, chronic inflammatory intestinal disease(Garcia Herola A. et al., Gastroenterol Hepatol. 2000 January; 23(1):16), celiac disease (Landau Y E. and Shoenfeld Y. Harefuah 2000 Jan.16; 138 (2):122), autoimmune diseases of the musculature, myositis,autoimmune myositis, Sjogren's syndrome (Feist E. et al., Int ArchAllergy Immunol 2000 September; 123 (1):92); smooth muscle autoimmunedisease (Zauli D. et al., Biomed Pharmacother 1999 June; 53 (5-6):234),hepatic diseases, hepatic autoimmune diseases, autoimmune hepatitis(Manns MP. J Hepatol 2000 August; 33 (2):326) and primary biliarycirrhosis (Strassburg C P. et al., Eur J Gastroenterol Hepatol. 1999June; 11 (6):595).

Type IV or T cell mediated hypersensitivity, include, but are notlimited to, rheumatoid diseases, rheumatoid arthritis (Tisch R, McDevittH O. Proc Natl Acad Sci USA 1994 Jan. 18; 91 (2):437), systemicdiseases, systemic autoimmune diseases, systemic lupus erythematosus(Datta S K., Lupus 1998; 7 (9):591), glandular diseases, glandularautoimmune diseases, pancreatic diseases, pancreatic autoimmunediseases, Type 1 diabetes (Castano L. and Eisenbarth G S. Ann. Rev.Immunol. 8:647); thyroid diseases, autoimmune thyroid diseases, Graves'disease (Sakata S. et al., Mol Cell Endocrinol 1993 March; 92 (1):77);ovarian diseases (Garza KM. et al., J Reprod Immunol 1998 February; 37(2):87), prostatitis, autoimmune prostatitis (Alexander R B. et al.,Urology 1997 December; 50 (6):893), polyglandular syndrome, autoimmunepolyglandular syndrome, Type I autoimmune polyglandular syndrome (HaraT. et al., Blood. 1991 Mar. 1; 77 (5):1127), neurological diseases,autoimmune neurological diseases, multiple sclerosis, neuritis, opticneuritis (Soderstrom M. et al., J Neurol Neurosurg Psychiatry 1994 May;57 (5):544), myasthenia gravis (Oshima M. et al., Eur J Immunol 1990December; 20 (12):2563), stiff-man syndrome (Hiemstra H S. et al., ProcNatl Acad Sci USA 2001 Mar. 27; 98 (7):3988), cardiovascular diseases,cardiac autoimmunity in Chagas' disease (Cunha-Neto E. et al., J ClinInvest 1996 Oct. 15; 98 (8):1709), autoimmune thrombocytopenic purpura(Semple J W. et al., Blood 1996 May 15; 87 (10):4245), anti-helper Tlymphocyte autoimmunity (Caporossi A P. et al., Viral Immunol 1998; 11(1):9), hemolytic anemia (Sallah S. et al., Ann Hematol 1997 March; 74(3):139), hepatic diseases, hepatic autoimmune diseases, hepatitis,chronic active hepatitis (Franco A. et al., Clin Immunol Immunopathol1990 March; 54 (3):382), biliary cirrhosis, primary biliary cirrhosis(Jones D E. Clin Sci (Colch) 1996 November; 91 (5):551), nephricdiseases, nephric autoimmune diseases, nephritis, interstitial nephritis(Kelly CJ. J Am Soc Nephrol 1990 August; 1 (2):140), connective tissuediseases, ear diseases, autoimmune connective tissue diseases,autoimmune ear disease (Yoo T J. et al., Cell Immunol 1994 August; 157(1):249), disease of the inner ear (Gloddek B. et al., Ann N Y Acad Sci1997 Dec. 29; 830:266), skin diseases, cutaneous diseases, dermaldiseases, bullous skin diseases, pemphigus vulgaris, bullous pemphigoidand pemphigus foliaceus.

Examples of delayed type hypersensitivity include, but are not limitedto, contact dermatitis and drug eruption.

Examples of types of T lymphocyte mediating hypersensitivity include,but are not limited to, helper T lymphocytes and cytotoxic Tlymphocytes.

Examples of helper T lymphocyte-mediated hypersensitivity include, butare not limited to, T_(h)1 lymphocyte mediated hypersensitivity andT_(h)2 lymphocyte mediated hypersensitivity.

According to a specific embodiment, the ocular disease is age-relatedmacular degeneration (AMD).

According to a specific embodiment, the age-related macular degeneration(AMD) is atrophic, non-neovascular (aAMD).

According to a specific embodiment, the age-related macular degeneration(AMD) is neovascular.

Autoimmune Diseases:

Autoimmune diseases include, but are not limited to, cardiovasculardiseases, rheumatoid diseases, glandular diseases, gastrointestinaldiseases, cutaneous diseases, hepatic diseases, neurological diseases,muscular diseases, nephric diseases, diseases related to reproduction,connective tissue diseases and systemic diseases.

Examples of autoimmune cardiovascular diseases include, but are notlimited to atherosclerosis (Matsuura E. et al., Lupus. 1998; 7 Suppl2:5135), myocardial infarction (Vaarala O. Lupus. 1998; 7 Suppl 2:S132),thrombosis (Tincani A. et al., Lupus 1998; 7 Suppl 2:S107-9), Wegener'sgranulomatosis, Takayasu's arteritis, Kawasaki syndrome (Praprotnik S.et al., Wien Klin Wochenschr 2000 Aug. 25; 112 (15-16):660), anti-factorVIII autoimmune disease (Lacroix-Desmazes S. et al., Semin ThrombHemost.2000; 26 (2):157), necrotizing small vessel vasculitis,microscopic polyangiitis, Churg and Strauss syndrome, pauci-immune focalnecrotizing and crescentic glomerulonephritis (Noel LH. Ann Med Interne(Paris). 2000 May; 151 (3):178), antiphospholipid syndrome (Flamholz R.et al., J Clin Apheresis 1999; 14 (4):171), antibody-induced heartfailure (Wallukat G. et al., Am J Cardiol. 1999 Jun. 17; 83 (12A):75H),thrombocytopenic purpura (Moccia F. Ann Ital Med Int. 1999 April-June;14 (2):114; Semple J W. et al., Blood 1996 May 15; 87 (10):4245),autoimmune hemolytic anemia (Efremov DG. et al., Leuk Lymphoma 1998January; 28 (3-4):285; Sallah S. et al., Ann Hematol 1997 March; 74(3):139), cardiac autoimmunity in Chagas' disease (Cunha-Neto E. et al.,J Clin Invest 1996 Oct. 15; 98 (8):1709) and anti-helper T lymphocyteautoimmunity (Caporossi AP. et al., Viral Immunol 1998; 11 (1):9).

Examples of autoimmune rheumatoid diseases include, but are not limitedto rheumatoid arthritis (Krenn V. et al., Histol Histopathol 2000 July;15 (3):791; Tisch R, McDevitt H O. Proc Natl Acad Sci units S A 1994Jan. 18; 91 (2):437) and ankylosing spondylitis (Jan Voswinkel et al.,Arthritis Res 2001; 3 (3): 189).

Examples of autoimmune glandular diseases include, but are not limitedto, pancreatic disease, Type I diabetes, thyroid disease, Graves'disease, thyroiditis, spontaneous autoimmune thyroiditis, Hashimoto'sthyroiditis, idiopathic myxedema, ovarian autoimmunity, autoimmuneanti-sperm infertility, autoimmune prostatitis and Type I autoimmunepolyglandular syndrome. diseases include, but are not limited toautoimmune diseases of the pancreas, Type 1 diabetes (Castano L. andEisenbarth GS. Ann. Rev. Immunol. 8:647; Zimmet P. Diabetes Res ClinPract 1996 October; 34 Suppl:S125), autoimmune thyroid diseases, Graves'disease (Orgiazzi J. Endocrinol Metab Clin North Am 2000 June; 29(2):339; Sakata S. et al., Mol Cell Endocrinol 1993 March; 92 (1):77),spontaneous autoimmune thyroiditis (Braley-Mullen H. and Yu S, J Immunol2000 Dec. 15; 165 (12):7262), Hashimoto's thyroiditis (Toyoda N. et al.,Nippon Rinsho 1999 August; 57 (8):1810), idiopathic myxedema (Mitsuma T.Nippon Rinsho. 1999 August; 57 (8):1759), ovarian autoimmunity (GarzaKM. et al., J Reprod Immunol 1998 February; 37 (2):87), autoimmuneanti-sperm infertility (Diekman AB. et al., Am J Reprod Immunol. 2000March; 43 (3):134), autoimmune prostatitis (Alexander RB. et al.,Urology 1997 December; 50 (6):893) and Type I autoimmune polyglandularsyndrome (Hara T. et al., Blood. 1991 Mar. 1; 77 (5):1127).

Examples of autoimmune gastrointestinal diseases include, but are notlimited to, chronic inflammatory intestinal diseases (Garcia Herola A.et al., Gastroenterol Hepatol. 2000 January; 23 (1):16), celiac disease(Landau Y E. and Shoenfeld Y. Harefuah 2000 Jan. 16; 138 (2):122),colitis, ileitis and Crohn's disease.

Examples of autoimmune cutaneous diseases include, but are not limitedto, autoimmune bullous skin diseases, such as, but are not limited to,pemphigus vulgaris, bullous pemphigoid and pemphigus foliaceus.

Examples of autoimmune hepatic diseases include, but are not limited to,hepatitis, autoimmune chronic active hepatitis (Franco A. et al., ClinImmunol Immunopathol 1990 March; 54 (3):382), primary biliary cirrhosis(Jones D E. Clin Sci (Colch) 1996 November; 91 (5):551; Strassburg CP.et al., Eur J Gastroenterol Hepatol. 1999 June; 11 (6):595) andautoimmune hepatitis (Manns MP. J Hepatol 2000 August; 33 (2):326).

Examples of autoimmune neurological diseases include, but are notlimited to, multiple sclerosis (Cross AH. et al., J Neuroimmunol 2001Jan. 1; 112 (1-2):1), Alzheimer's disease (Oron L. et al., J NeuralTransm Suppl. 1997; 49:77), myasthenia gravis (Infante AJ. And Kraig E,Int Rev Immunol 1999; 18 (1-2):83; Oshima M. et al., Eur J Immunol 1990December; 20 (12):2563), neuropathies, motor neuropathies (Kornberg AJ.J Clin Neurosci. 2000 May; 7 (3):191); Guillain-Barre syndrome andautoimmune neuropathies (Kusunoki S. Am J Med Sci. 2000 April; 319(4):234), myasthenia, Lambert-Eaton myasthenic syndrome (Takamori M. AmJ Med Sci. 2000 April; 319 (4):204); paraneoplastic neurologicaldiseases, cerebellar atrophy, paraneoplastic cerebellar atrophy andstiff-man syndrome (Hiemstra H S. et al., Proc Natl Acad Sci units S A2001 Mar. 27; 98 (7):3988); non-paraneoplastic stiff man syndrome,progressive cerebellar atrophies, encephalitis, Rasmussen'sencephalitis, amyotrophic lateral sclerosis, Sydeham chorea, Gilles dela Tourette syndrome and autoimmune polyendocrinopathies (Antoine J C.and Honnorat J. Rev Neurol (Paris) 2000 January; 156 (1):23); dysimmuneneuropathies (Nobile-Orazio E. et al., Electroencephalogr ClinNeurophysiol Suppl 1999; 50:419); acquired neuromyotonia, arthrogryposismultiplex congenita (Vincent A. et al., Ann N Y Acad Sci. 1998 May 13;841:482), neuritis, optic neuritis (Soderstrom M. et al., J NeurolNeurosurg Psychiatry 1994 May; 57 (5):544) and neurodegenerativediseases.

Examples of autoimmune muscular diseases include, but are not limitedto, myositis, autoimmune myositis and primary Sjogren's syndrome (FeistE. et al., Int Arch Allergy Immunol 2000 September; 123 (1):92) andsmooth muscle autoimmune disease (Zauli D. et al., Biomed Pharmacother1999 June; 53 (5-6):234).

Examples of autoimmune nephric diseases include, but are not limited to,nephritis and autoimmune interstitial nephritis (Kelly CJ. J Am SocNephrol 1990 August; 1 (2):140).

Examples of autoimmune diseases related to reproduction include, but arenot limited to, repeated fetal loss (Tincani A. et al., Lupus 1998; 7Suppl 2:S107-9).

Examples of autoimmune connective tissue diseases include, but are notlimited to, ear diseases, autoimmune ear diseases (Yoo TJ. et al., CellImmunol 1994 August; 157 (1):249) and autoimmune diseases of the innerear (Gloddek B. et al., Ann N Y Acad Sci 1997 Dec. 29; 830:266).

Examples of autoimmune systemic diseases include, but are not limitedto, systemic lupus erythematosus (Erikson J. et al., Immunol Res 1998;17 (1-2):49) and systemic sclerosis (Renaudineau Y. et al., Clin DiagnLab Immunol. 1999 March; 6 (2):156); Chan OT. et al., Immunol Rev 1999June; 169:107).

According to one embodiment, the autoimmune disease is Crohn's disease,psoriasis, scleroderma or rheumatoid arthritis.

Graft Rejection Diseases:

Examples of diseases associated with transplantation of a graft include,but are not limited to, graft rejection, chronic graft rejection,subacute graft rejection, hyperacute graft rejection, acute graftrejection and graft versus host disease.

Allergic Diseases:

Examples of allergic diseases include, but are not limited to, asthma,hives, urticaria, pollen allergy, dust mite allergy, venom allergy,cosmetics allergy, latex allergy, chemical allergy, drug allergy, insectbite allergy, animal dander allergy, stinging plant allergy, poison ivyallergy and food allergy.

Pharmaceutical Compositions:

The compounds described herein according to any of the aspects ofembodiments of the invention described herein can be utilized (e.g.,administered to a subject) per se or in a pharmaceutical compositionwhere the compound is mixed with suitable carriers or excipients.

As used herein a “pharmaceutical composition” refers to a preparation ofone or more compound according to any of the embodiments describedherein with other chemical components such as physiologically suitablecarriers and excipients. The purpose of a pharmaceutical composition isto facilitate administration of a compound to an organism.

Hereinafter, the phrases “physiologically acceptable carrier” and“pharmaceutically acceptable carrier” which may be interchangeably usedrefer to a carrier or a diluent that does not cause significantirritation to an organism and does not abrogate the biological activityand properties of the administered compound. An adjuvant is includedunder these phrases.

Herein the term “excipient” refers to an inert substance added to apharmaceutical composition to further facilitate administration of anactive ingredient. Examples, without limitation, of excipients includecalcium carbonate, calcium phosphate, various sugars and types ofstarch, cellulose derivatives, gelatin, vegetable oils and polyethyleneglycols.

When utilized per se or in a pharmaceutically acceptable composition,the compound per se (that is, not including, weight of carriers orexcipients co-formulated with the compound, as described herein) isoptionally at least 80% pure (by dry weight), optionally at least 90%pure (by dry weight), at least 95% pure (by dry weight), at least 98%pure (by dry weight), and optionally at least 99% pure (by dry weight).Purity may be enhanced, e.g., by removing impurities associated withsynthesis of the compound or isolation of the compound from a naturalsource, by any suitable technique known in the art. As exemplifiedherein, impurities of a compound described herein (for example, BKT300)may weaken a biological effect of the compound.

Techniques for formulation and administration of drugs may be found in“Remington's Pharmaceutical Sciences”, Mack Publishing Co., Easton, Pa.,latest edition, which is incorporated herein by reference.

Suitable routes of administration may, for example, include oral,rectal, transmucosal, especially transnasal, intestinal or parenteraldelivery, including intramuscular, subcutaneous and intramedullaryinjections as well as intrathecal, direct intraventricular,intracardiac, e.g., into the right or left ventricular cavity, into thecommon coronary artery, intravenous, intraperitoneal, intranasal, orintraocular injections.

Alternately, one may administer the pharmaceutical composition in alocal rather than systemic manner, for example, via injection of thepharmaceutical composition directly into a tissue region of a patient.

The term “tissue” refers to part of an organism consisting of cellsdesigned to perform a function or functions. Examples include, but arenot limited to, brain tissue, retina, skin tissue, hepatic tissue,pancreatic tissue, breast tissue, bone, cartilage, connective tissue,blood tissue, muscle tissue, cardiac tissue brain tissue, vasculartissue, renal tissue, pulmonary tissue, gonadal tissue, hematopoietictissue.

Pharmaceutical compositions of some embodiments of the invention may bemanufactured by processes well known in the art, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or lyophilizing processes.

Pharmaceutical compositions for use in accordance with some embodimentsof the invention thus may be formulated in conventional manner using oneor more physiologically acceptable carriers comprising excipients andauxiliaries, which facilitate processing of the active ingredients intopreparations which, can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen.

For injection, the active ingredients of the pharmaceutical compositionmay be formulated in aqueous solutions, preferably in physiologicallycompatible buffers such as Hank's solution, Ringer's solution, orphysiological salt buffer. For transmucosal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art.

For oral administration, the pharmaceutical composition can beformulated readily by combining the active compounds withpharmaceutically acceptable carriers well known in the art. Suchcarriers enable the pharmaceutical composition to be formulated astablets, pills, dragees, capsules, liquids, gels, syrups, slurries,suspensions, and the like, for oral ingestion by a patient.Pharmacological preparations for oral use can be made using a solidexcipient, optionally grinding the resulting mixture, and processing themixture of granules, after adding suitable auxiliaries if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose; and/or physiologically acceptable polymers suchas polyvinylpyrrolidone (PVP). If desired, disintegrating agents may beadded, such as cross-linked polyvinyl pyrrolidone, agar, or alginic acidor a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, titanium dioxide, lacquer solutions and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Pharmaceutical compositions which can be used orally include push-fitcapsules made of gelatin as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules may contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, lubricants such as talc ormagnesium stearate and, optionally, stabilizers. In soft capsules, theactive ingredients may be dissolved or suspended in suitable liquids,such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers may be added. All formulations for oraladministration should be in dosages suitable for the chosen route ofadministration.

For buccal administration, the compositions may take the form of tabletsor lozenges formulated in conventional manner.

For administration by nasal inhalation, the active ingredients for useaccording to some embodiments of the invention are convenientlydelivered in the form of an aerosol spray presentation from apressurized pack or a nebulizer with the use of a suitable propellant,e.g., dichlorodifluoromethane, trichlorofluoromethane,dichloro-tetrafluoroethane or carbon dioxide. In the case of apressurized aerosol, the dosage unit may be determined by providing avalve to deliver a metered amount. Capsules and cartridges of, e.g.,gelatin for use in a dispenser may be formulated containing a powder mixof the active compound and a suitable powder base such as lactose orstarch.

The pharmaceutical composition described herein may be formulated forparenteral administration, e.g., by bolus injection or continuousinfusion. Formulations for injection may be presented in unit dosageform, e.g., in ampoules or in multidose containers with optionally, anadded preservative. The compositions may be suspensions, solutions oremulsions in oily or aqueous vehicles, and may contain formulatoryagents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical compositions for parenteral administration includeaqueous solutions of the active preparation in water-soluble form.Additionally, suspensions of the active ingredients may be prepared asappropriate oily or water based injection suspensions. Suitablelipophilic solvents or vehicles include fatty oils such as sesame oil,or synthetic fatty acids esters such as ethyl oleate, triglycerides orliposomes.

Aqueous injection suspensions may contain substances, which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol or dextran.

Optionally, the suspension may also contain suitable stabilizers oragents which increase the solubility of the active ingredients to allowfor the preparation of highly concentrated solutions.

Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile, pyrogen-free waterbased solution, before use.

The pharmaceutical composition of some embodiments of the invention mayalso be formulated in rectal compositions such as suppositories orretention enemas, using, e.g., conventional suppository bases such ascocoa butter or other glycerides.

Pharmaceutical compositions suitable for use in context of someembodiments of the invention include compositions wherein the activeingredients are contained in an amount effective to achieve the intendedpurpose. More specifically, a therapeutically effective amount means anamount of the active ingredient(s) effective to prevent, alleviate orameliorate symptoms of a disorder (e.g., cancer or metastatic cancer) orprolong the survival of the subject being treated.

Determination of a therapeutically effective amount is well within thecapability of those skilled in the art, especially in light of thedetailed disclosure provided herein.

For any preparation used in the methods of the invention, thetherapeutically effective amount or dose can be estimated initially fromin vitro and cell culture assays. For example, a dose can be formulatedin animal models to achieve a desired concentration or titer. Suchinformation can be used to more accurately determine useful doses inhumans.

Toxicity and therapeutic efficacy of the active ingredients describedherein can be determined by standard pharmaceutical procedures in vitro,in cell cultures or experimental animals. The data obtained from thesein vitro and cell culture assays and animal studies can be used informulating a range of dosage for use in human. The dosage may varydepending upon the dosage form employed and the route of administrationutilized. The exact formulation, route of administration and dosage canbe chosen by the individual physician in view of the patient's condition(see, e.g., Fingl et al. (1975), in “The Pharmacological Basis ofTherapeutics”, Ch. 1 p. 1).

Dosage amount and interval may be adjusted individually to provideprotein (e.g., MIP3a, MCP-1, SDF-1 and/or CXCR4, and/or a kinase)inhibitory levels of the active ingredient which are sufficient toinduce or suppress the biological effect (minimal effectiveconcentration, MEC). The MEC will vary for each preparation, but can beestimated from in vitro data, e.g., based on results of achemokine-induced (e.g., MIP3a-, MCP-1- and/or SDF-1-induced) migrationinhibition assay described herein and/or results of a kinase inhibitionassay and/or results of cell viability assay as described herein.Dosages necessary to achieve the MEC will depend on individualcharacteristics and route of administration. Detection assays can beused to determine plasma concentrations.

In some embodiments of any of the embodiments described herein, aneffective amount of the compound is less than 100 μM. In someembodiments, an effective amount is less than 10 μM. In someembodiments, an effective amount is less than 5 μM. In some embodiments,an effective amount is less than 2.5 μM.

In some embodiments of any of the embodiments described herein, aneffective amount is at least 100% of the IC50 of the compound towards achemokine which is intended to be inhibited (e.g., MIP3a, MCP-1 and/orSDF-1). In some embodiments, an effective amount is at least 200% of theIC50 of the compound towards the chemokine.

In some embodiments, an effective amount is at least 300% of the IC50 ofthe compound towards the chemokine. In some embodiments, an effectiveamount is at least 500% of the IC50 of the compound towards thechemokine. In some embodiments, an effective amount is at least 1000% ofthe IC50 of the compound towards the chemokine.

In some embodiments of any of the embodiments described herein, aneffective amount is at least 100% of the IC50 of the compound towardsinducing cell death of cancer cells to be inhibited. In someembodiments, an effective amount is at least 200% of the IC50 of thecompound towards the cancer cells. In some embodiments, an effectiveamount is at least 300% of the IC50 of the compound towards the cancercells.

In some embodiments of any of the embodiments described herein, aneffective amount is at least 100% of the IC50 of the compound towards akinase. In some embodiments, an effective amount is at least 200% of theIC50 of the compound towards a kinase. In some embodiments, an effectiveamount is at least 300% of the IC50 of the compound towards the kinase.

Depending on the severity and responsiveness of the condition to betreated, dosing can be of a single or a plurality of administrations,with course of treatment lasting from several days to several weeks oruntil cure is effected or diminution of the disease state is achieved.

The amount of a composition to be administered will, of course, bedependent on the subject being treated, the severity of the affliction,the manner of administration, the judgment of the prescribing physician,etc.

Compositions of some embodiments of the invention may, if desired, bepresented in a pack or dispenser device, such as an FDA approved kit,which may contain one or more unit dosage forms containing the activeingredient. The pack may, for example, comprise metal or plastic foil,such as a blister pack. The pack or dispenser device may be accompaniedby instructions for administration. The pack or dispenser may also beaccommodated by a notice associated with the container in a formprescribed by a governmental agency regulating the manufacture, use orsale of pharmaceuticals, which notice is reflective of approval by theagency of the form of the compositions or human or veterinaryadministration. Such notice, for example, may be of labeling approved bythe U.S. Food and Drug Administration for prescription drugs or of anapproved product insert. Compositions comprising a preparation of theinvention formulated in a compatible pharmaceutical carrier may also beprepared, placed in an appropriate container, and labeled for treatmentof an indicated condition, as is further detailed herein.

According to another aspect described herein, there is provided a kitfor the treatment of a condition (e.g., treatment of cancer orprevention of tumor metastasis or treatment of non-cancerousproliferative disease or disorder or treatment of inflammation)described herein, the kit comprising a packaging material packaging thecompound described herein.

In some embodiments, the compound is identified as an inhibitor of anSDF-1 and/or CXCR4 activity associated with an onset or progression ofthe condition, as described herein.

In some embodiments, the compound is identified as an inhibitor of akinase activity associated with an onset or progression of thecondition, as described herein.

In some embodiments, the compound is identified as inducing apoptosisand/or cell growth arrest of cells associated with the condition, asdescribed herein.

It will be appreciated that the compounds described herein can beprovided or utilized, in any of the methods, uses, compositions and kitsdescribed herein, and any embodiments thereof, alone or in combinationwith other active ingredients, which are well known in the art foralleviating the medical condition.

Thus, for example, the compound may be administered with animmunomodulator, either together in a co-formulation or in separateformulations.

According to a specific embodiment, the treatment of cancer (and otherhyperproliferative disorders) is effected in combination with ananti-cancer immune modulator agent.

As used herein, the term “anti-cancer immune modulator agent” refers toan agent capable of eliciting an immune response (e.g. T cell, NK cell)against a cancerous cell.

According to specific embodiment, the agent is selected from the groupconsisting of a cancer antigen, a cancer vaccine, an anti-cancerantibody, a cytokine capable of inducing activation and/or proliferationof a T cell and an immune-check point regulator.

Alternatively or additionally, such modulators may be immune stimulatorssuch as immune-check point regulators which are of specific value in thetreatment of cancer.

As used herein the term “immune-check point regulator” refers to amolecule that modulates the activity of one or more immune-check pointproteins in an agonistic or antagonistic manner resulting in activationof an immune cell.

As used herein the term “immune-check point protein” refers to a proteinthat regulates an immune cell activation or function. Immune check-pointproteins can be either co-stimulatory proteins (i.e. transmitting astimulatory signal resulting in activation of an immune cell) orinhibitory proteins (i.e. transmitting an inhibitory signal resulting insuppressing activity of an immune cell). According to specificembodiment, the immune check point protein regulates activation orfunction of a T cell. Numerous checkpoint proteins are known in the artand include, but not limited to, PD1, PDL-1, B7H2, B7H4, CTLA-4, CD80,CD86, LAG-3, TIM-3, KIR, IDO, CD19, OX40, 4-1BB (CD137), CD27, CD70,CD40, GITR, CD28 and ICOS (CD278).

According to specific embodiments, the immune-check-point regulator isselected form the group consisting of anti-CTLA4, anti-PD-1, and CD40agonist.

According to specific embodiments, the immune-check point regulator isselected form the group consisting of anti-CTLA4, anti-PD-1, anti-PDL-1,CD40 agonist, 4-1BB agonist, GITR agonist and OX40 agonist.

CTLA4 is a member of the immunoglobulin superfamily, which is expressedon the surface of Helper T cells and transmits an inhibitory signal to Tcells upon ligand binding. As used herein, the term “anti-CTLA4” refersto an antagonistic molecule that binds CTLA4 (CD152) and suppresses itssuppressive activity. Thus, an anti-CTLA4 prevents the transmission ofthe inhibitory signal and thereby acts as a co-stimulatory molecule.According to a specific embodiment, the anti-CDLA4 molecule is anantibody.

PD-1 (Programmed Death 1) is a member of the extended CD28/CTLA-4 familyof T cell regulators which is expressed on the surface of activated Tcells, B cells, and macrophages and transmits an inhibitory signal uponligand binding. As used herein, the term “anti-PD1” refers to anantagonistic molecule that binds PD-1 and suppresses its suppressiveactivity. Thus, an anti-PD-1 prevents the transmission of the inhibitorysignal and thereby acts as a co-stimulatory molecule. According to aspecific embodiment, the anti-PD1 molecule is an antibody. Numerousanti-PD-1 antibodies are known in the art see e.g. Topalian, et al. NEJM2012.

PDL-1 is a ligand of PD-1. Binding of PDL-1 to its receptor PD-1transmits an inhibitory signal to the cell expressing the PD-1. As usedherein, the tem “anti-PDL-1” refers to an antagonistic molecule thatinhibits PD-1 signaling by binding to or inhibiting PD-L1 from bindingand/or activating PD-1. Thus, an anti-PD-1 prevents the transmission ofthe inhibitory signal and thereby acts as a co-stimulatory molecule.According to specific embodiments, the anti-PD-L1 is an anti-PD-L1antibody. Numerous anti-PDL-1 antibodies are known in the art see e.g.Brahmer, et al. NEJM 2012.

CD40 (CD154) is a co-stimulatory receptor found on antigen presentingcells and transmits an activation signal upon ligand binding. As usedherein, the term “CD40 agonist” refers to an agonistic molecule thatbinds CD40 (CD154) and thereby induces activation of the antigenpresenting cell.

OX40 belongs to the TNF receptor super family and leads to expansion ofCD4+ and CD8+ T cells. As used herein, the term “OX40 agonist” refers toan agonistic molecule that binds and activates OX40.

GITR (glucocorticoid-induced tumor necrosis factor receptor) is asurface receptor molecule that has been shown to be involved ininhibiting the suppressive activity of T-regulatory cells and extendingthe survival of T-effector cells. As used herein, the term “GITRagonist” refers to an agonistic molecule that binds and activates GITR.According to a specific embodiment, the GITR agonist is an antibody.

Definitions

As used herein, the term “treating” includes abrogating, substantiallyinhibiting, slowing or reversing the progression of a condition,substantially ameliorating clinical or aesthetical symptoms of acondition or substantially preventing the appearance of clinical oraesthetical symptoms of a condition. For example, in the context ofpreventing metastasis and/or angiogenesis, the term “preventing” refersto arresting, halting, inhibiting the metastatic and/or angiogeneticprocess or progression and subsequent metastasis and/or angiogenesis.

As used herein the term “subject” refers to a mammal (e.g., human), forexample, one who has been diagnosed with a condition described herein(e.g., cancer).

The terms “comprises”, “comprising”, “includes”, “including”, “having”and their conjugates mean “including but not limited to”.

The term “consisting of” means “including and limited to”.

The term “consisting essentially of” means that the composition, methodor structure may include additional ingredients, steps and/or parts, butonly if the additional ingredients, steps and/or parts do not materiallyalter the basic and novel characteristics of the claimed composition,method or structure.

As used herein, the singular form “a”, “an” and “the” include pluralreferences unless the context clearly dictates otherwise. For example,the term “a compound” or “at least one compound” may include a pluralityof compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention maybe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 3, 4, 5, and 6. This appliesregardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to includeany cited numeral (fractional or integral) within the indicated range.The phrases “ranging/ranges between” a first indicate number and asecond indicate number and “ranging/ranges from” a first indicate number“to” a second indicate number are used herein interchangeably and aremeant to include the first and second indicated numbers and all thefractional and integral numerals therebetween.

As used herein the term “method” refers to manners, means, techniquesand procedures for accomplishing a given task including, but not limitedto, those manners, means, techniques and procedures either known to, orreadily developed from known manners, means, techniques and proceduresby practitioners of the chemical, pharmacological, biological,biochemical and medical arts.

Herein throughout, the phrase “linking group” describes a group (asubstituent) that is attached to another moiety in the compound via twoor more atoms thereof. In order to differentiate a linking group from asubstituent that is attached to another moiety in the compound via oneatom thereof, the latter will be referred to herein and throughout as an“end group”.

As used herein, the term “amine” describes both a —NR′R″ end group and a—NR′— linking group, wherein R′ and R″ are each independently hydrogen,alkyl, cycloalkyl, aryl, as these terms are defined hereinbelow.

The amine group can therefore be a primary amine, where both R′ and R″are hydrogen, a secondary amine, where R′ is hydrogen and R″ is alkyl,cycloalkyl or aryl, or a tertiary amine, where each of R′ and R″ isindependently alkyl, cycloalkyl or aryl.

Alternatively, R′ and R″ can each independently be hydroxyalkyl,trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl,heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate,hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano,nitro, azo, sulfonamide, carbonyl, C-carboxylate, O-carboxylate,N-thiocarbamate, O-thiocarbamate, urea, thiourea, N-carbamate,O-carbamate, C-amide, N-amide, guanyl, guanidine and hydrazine.

The term “amine” is used herein to describe a —NR′R″ group in caseswhere the amine is an end group, as defined hereinunder, and is usedherein to describe a —NR′— group in cases where the amine is or forms apart of a linking group.

The term “alkyl” describes a saturated aliphatic hydrocarbon includingstraight chain and branched chain groups. Preferably, the alkyl grouphas 1 to 20 carbon atoms. Whenever a numerical range; e.g., “1-20”, isstated herein, it implies that the group, in this case the alkyl group,may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up toand including 20 carbon atoms. In some embodiments, the alkyl is amedium size alkyl having 1 to 10 carbon atoms. Unless otherwiseindicated, the alkyl is a lower alkyl having 1 to 4 carbon atoms. Insome embodiments, the alkyl has at least 4 carbon atoms, for example,the alkyl is having 4 to 12 or 4 to 10 or 4 to 8 carbon atoms. The alkylgroup may be substituted or unsubstituted. Substituted alkyl may haveone or more substituents, whereby each substituent group canindependently be, for example, hydroxyalkyl, trihaloalkyl, cycloalkyl,alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide,sulfinate, sulfate, sulfonate, sulfoxide, phosphonate, oxo, hydroxy,alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro,azo, sulfonamide, C-carboxylate, O-carboxylate, N-thiocarbamate,O-thiocarbamate, urea, thiourea, N-carbamate, O-carbamate, C-amide,N-amide, guanyl, guanidine and hydrazine.

The alkyl group can be an end group, as this phrase is definedhereinabove, wherein it is attached to a single adjacent atom, or alinking group, as this phrase is defined hereinabove, which connects twoor more moieties via at least two carbons in its chain. When an alkyl isa linking group, it is also referred to herein as “alkylene”, e.g.,methylene, ethylene, propylene, etc.

The term “alkenyl” describes an alkyl, as defined herein, in which atleast one pair of carbon atoms are linked to one another via a doublebond.

The term “alkynyl” or “alkyne” describes an alkyl, as defined herein, inwhich at least one pair of carbon atoms are linked to one another via atriple bond.

The term “cycloalkyl” describes an all-carbon monocyclic or fused ring(i.e., rings which share an adjacent pair of carbon atoms) group whereone or more of the rings does not have a completely conjugatedpi-electron system. The cycloalkyl group may be substituted orunsubstituted. Substituted cycloalkyl may have one or more substituents,whereby each substituent group can independently be, for example,hydroxyalkyl, trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl,heteroaryl, heteroalicyclic, amine, halide, oxo, sulfinate, sulfate,sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide,C-carboxylate, O-carboxylate, N-thiocarbamate, O-thiocarbamate, urea,thiourea, N-carbamate, O-carbamate, C-amide, N-amide, guanyl, guanidineand hydrazine. The cycloalkyl group can be an end group, as this phraseis defined hereinabove, wherein it is attached to a single adjacentatom, or a linking group, as this phrase is defined hereinabove,connecting two or more moieties at two or more positions thereof.

The term “heteroalicyclic” describes a monocyclic or fused ring grouphaving in the ring(s) one or more atoms such as nitrogen, oxygen andsulfur. The rings may also have one or more double bonds. However, therings do not have a completely conjugated pi-electron system. Theheteroalicyclic may be substituted or unsubstituted.

Substituted heteroalicyclic may have one or more substituents, wherebyeach substituent group can independently be, for example, hydroxyalkyl,trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl,heteroalicyclic, amine, halide, oxo, sulfinate, sulfate, sulfonate,sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy,thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, C-carboxylate,O-carboxylate, N-thiocarbamate, O-thiocarbamate, urea, thiourea,O-carbamate, N-carbamate, C-amide, N-amide, guanyl, guanidine andhydrazine. The heteroalicyclic group can be an end group, as this phraseis defined hereinabove, where it is attached to a single adjacent atom,or a linking group, as this phrase is defined hereinabove, connectingtwo or more moieties at two or more positions thereof. Representativeexamples are piperidine, piperazine, tetrahydrofurane, tetrahydropyrane,morpholino and the like.

The term “aryl” describes an all-carbon monocyclic or fused-ringpolycyclic (i.e., rings which share adjacent pairs of carbon atoms)groups having a completely conjugated pi-electron system. The aryl groupmay be substituted or unsubstituted.

Substituted aryl may have one or more substituents, whereby eachsubstituent group can independently be, for example, hydroxyalkyl,trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl,heteroalicyclic, amine, halide, oxo, sulfinate, sulfate, sulfonate,sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy,thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, C-carboxylate,O-carboxylate, N-thiocarbamate, O-thiocarbamate, urea, thiourea,N-carbamate, O-carbamate, C-amide, N-amide, guanyl, guanidine andhydrazine. The aryl group can be an end group, as this term is definedhereinabove, wherein it is attached to a single adjacent atom, or alinking group, as this term is defined hereinabove, connecting two ormore moieties at two or more positions thereof. Preferably, the aryl isphenyl. Optionally, the aryl is naphthalenyl.

The term “heteroaryl” describes a monocyclic or fused ring (i.e., ringswhich share an adjacent pair of atoms) group having in the ring(s) oneor more atoms, such as, for example, nitrogen, oxygen and sulfur and, inaddition, having a completely conjugated pi-electron system. Examples,without limitation, of heteroaryl groups include pyrrole, furane,thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine,triazine, tetrazine, quinoline, isoquinoline and purine. The heteroarylgroup may be substituted or unsubstituted. Substituted heteroaryl mayhave one or more substituents, whereby each substituent group canindependently be, for example, hydroxyalkyl, trihaloalkyl, cycloalkyl,alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide,sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide,C-carboxylate, O-carboxylate, N-thiocarbamate, O-thiocarbamate, urea,thiourea, O-carbamate, N-carbamate, C-amide, N-amide, guanyl, guanidineand hydrazine. The heteroaryl group can be an end group, as this phraseis defined hereinabove, where it is attached to a single adjacent atom,or a linking group, as this phrase is defined hereinabove, connectingtwo or more moieties at two or more positions thereof.

The term “alkaryl” describes an alkyl, as defined herein, which issubstituted by one or more aryl or heteroaryl groups. An example ofalkaryl is benzyl.

The term “halide” and “halo” describes fluorine, chlorine, bromine oriodine.

The term “haloalkyl” describes an alkyl group as defined above, furthersubstituted by one or more halide.

The term “sulfate” describes a —O—S(═O)₂—OR′ end group, as this term isdefined hereinabove, or an —O—S(═O)₂—O— linking group, as these phrasesare defined hereinabove, where R′ is as defined hereinabove.

The term “thiosulfate” describes a —O—S(═S)(═O)—OR′ end group or a—O—S(═S)(═O)—O— linking group, as these phrases are defined hereinabove,where R′ is as defined hereinabove.

The term “sulfite” describes an —O—S(═O)—O—R′ end group or a —O—S(═O)—O—group linking group, as these phrases are defined hereinabove, where R′is as defined hereinabove.

The term “thiosulfite” describes a —O—S(═S)—O—R′ end group or an—O—S(═S)—O— group linking group, as these phrases are definedhereinabove, where R′ is as defined hereinabove.

The term “sulfinate” or “sulfinyl” describes a —S(═O)—OR′ end group oran —S(═O)—O— group linking group, as these phrases are definedhereinabove, where R′ is as defined hereinabove.

The term “sulfoxide” describes a —S(═O)R′ end group or an —S(═O)—linking group, as these phrases are defined hereinabove, where R′ is asdefined hereinabove.

The term “sulfonate” or “sulfonyl” describes a —S(═O)₂—OR′ end group(also referred to herein as —SO₃R′ or —SO₃H) or an —O—S(═O)₂— linkinggroup, as these phrases are defined hereinabove, where R′ is as definedherein.

The term “S-sulfonamide” describes a —S(═O)₂—NR′R″ end group or a—S(═O)₂—NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “N-sulfonamide” describes an R'S(═O)₂—NR″— end group or a—S(═O)₂—NR′— linking group, as these phrases are defined hereinabove,where R′ and R″ are as defined herein.

The term “disulfide” refers to a —S—SR′ end group or a —S—S— linkinggroup, as these phrases are defined hereinabove, where R′ is as definedherein.

The term “phosphonate” describes a —P(═O)(OR′)(OR″) end group or a—P(═O)(OR′)(O)— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “thiophosphonate” describes a —P(═S)(OR′)(OR″) end group or a—P(═S)(OR′)(O)— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “carbonyl” or “carbonate” as used herein, describes a —C(═O)—R′end group or a —C(═O)— linking group, as these phrases are definedhereinabove, with R′ as defined herein.

The term “thiocarbonyl” as used herein, describes a —C(═S)—R′ end groupor a —C(═S)— linking group, as these phrases are defined hereinabove,with R′ as defined herein.

The term “oxo” as used herein, described a ═O end group.

The term “thiooxo” as used herein, described a ═S end group.

The term “oxime” describes a ═N—OH end group or a ═N—O— linking group,as these phrases are defined hereinabove.

The term “hydroxyl” or “hydroxy” describes a —OH group.

The term “alkoxy” describes both an —O-alkyl and an —O-cycloalkyl group,as defined herein.

The term “aryloxy” describes both an —O-aryl and an —O-heteroaryl group,as defined herein.

The term “thiohydroxy” or “thio” describes a —SH group.

The term “thioalkoxy” describes both a —S-alkyl group, and a—S-cycloalkyl group, as defined herein.

The term “thioaryloxy” describes both a —S-aryl and a —S-heteroarylgroup, as defined herein.

The term “cyano” or “nitrile” describes a —C≡N group.

The term “isocyanate” describes an —N═C═O group.

The term “nitro” describes an —NO₂ group.

The term “carboxylate” as used herein encompasses C-carboxylate andO-carboxylate.

The term “C-carboxylate” describes a —C(═O)—OR′ end group or a —C(═O)—O—linking group, as these phrases are defined hereinabove, where R′ is asdefined herein.

The term “O-carboxylate” describes a —OC(═O)R′ end group or a—OC(═O)-linking group, as these phrases are defined hereinabove, whereR′ is as defined herein.

The term “thiocarboxylate” as used herein encompasses “C-thiocarboxylateand O-thiocarboxylate.

The term “C-thiocarboxylate” describes a —C(═S)—OR′ end group or a—C(═S)—O— linking group, as these phrases are defined hereinabove, whereR′ is as defined herein.

The term “O-thiocarboxylate” describes a —OC(═S)R′ end group or a—OC(═S)-linking group, as these phrases are defined hereinabove, whereR′ is as defined herein.

The term “carbamate” as used herein encompasses N-carbamate andO-carbamate.

The term “N-carbamate” describes an R″OC(═O)—NR′— end group or a—OC(═O)—NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “O-carbamate” describes an —OC(═O)—NR′R″ end group or an—OC(═O)—NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “thiocarbamate” as used herein encompasses N-thiocarbamate andO-thiocarbamate.

The term “O-thiocarbamate” describes a —OC(═S)—NR′R″ end group or a—OC(═S)—NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “N-thiocarbamate” describes an R″OC(═S)NR′— end group or a—OC(═S)NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “dithiocarbamate” as used herein encompasses N-dithiocarbamateand S-dithiocarbamate.

The term “S-dithiocarbamate” describes a —SC(═S)—NR′R″ end group or a—SC(═S)NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “N-dithiocarbamate” describes an R″SC(═S)NR′— end group or a—SC(═S)NR′— linking group, as these phrases are defined hereinabove,with R′ and R″ as defined herein.

The term “urea”, which is also referred to herein as “ureido”, describesa —NR′C(═O)—NR″R′″ end group or a —NR′C(═O)—NR″— linking group, as thesephrases are defined hereinabove, where R′ and R″ are as defined hereinand R′″ is as defined herein for R′ and R″.

The term “thiourea”, which is also referred to herein as “thioureido”,describes a —NR′—C(═S)—NR″R′″ end group or a —NR′—C(═S)—NR″— linkinggroup, with R′, R″ and R′″ as defined herein.

The term “amide” as used herein encompasses C-amide and N-amide.

The term “C-amide” describes a —C(═O)—NR′R″ end group or a—C(═O)—NR′-linking group, as these phrases are defined hereinabove,where R′ and R″ are as defined herein.

The term “N-amide” describes a R′C(═O)—NR″— end group or a R′C(═O)—N—linking group, as these phrases are defined hereinabove, where R′ and R″are as defined herein.

The term “guanyl” describes a R′R″NC(═N)— end group or a—R′NC(═N)-linking group, as these phrases are defined hereinabove, whereR′ and R″ are as defined herein.

The term “guanidine” describes a —R′NC(═N)—NR″R′″ end group or a—R′NC(═N)— NR″— linking group, as these phrases are defined hereinabove,where R′, R″ and R′″ are as defined herein.

The term “hydrazine” describes a —NR′—NR″R′″ end group or a—NR′—NR″-linking group, as these phrases are defined hereinabove, withR′, R″, and R′″ as defined herein.

As used herein, the term “hydrazide” describes a —C(═O)—NR′—NR″R′″ endgroup or a —C(═O)—NR′—NR″— linking group, as these phrases are definedhereinabove, where R′, R″ and R′″ are as defined herein.

As used herein, the term “thiohydrazide” describes a —C(═S)—NR′—NR″R′″end group or a —C(═S)—NR′—NR″— linking group, as these phrases aredefined hereinabove, where R′, R″ and R′″ are as defined herein.

For any of the embodiments described herein, the compound describedherein may be in a form of a salt, for example, a pharmaceuticallyacceptable salt, and/or in a form of a prodrug.

As used herein, the phrase “pharmaceutically acceptable salt” refers toa charged species of the parent compound and its counter-ion, which istypically used to modify the solubility characteristics of the parentcompound and/or to reduce any significant irritation to an organism bythe parent compound, while not abrogating the biological activity andproperties of the administered compound.

In the context of some of the present embodiments, a pharmaceuticallyacceptable salt of the compounds described herein may optionally be abase addition salt comprising at least one acidic (e.g., phenol and/orcarboxylic acid) group of the compound which is in a negatively chargedform (e.g., wherein the acidic group is deprotonated), in combinationwith at least one counter-ion, derived from the selected base, thatforms a pharmaceutically acceptable salt.

The base addition salts of the compounds described herein may thereforebe complexes formed between one or more acidic groups of the drug andone or more equivalents of a base.

The base addition salts may include a variety of organic and inorganiccounter-ions and bases, such as, but not limited to, sodium (e.g., byaddition of NaOH), potassium (e.g., by addition of KOH), calcium (e.g.,by addition of Ca(OH)₂, magnesium (e.g., by addition of Mg(OH)₂),aluminum (e.g., by addition of Al(OH)₃ and ammonium (e.g., by additionof ammonia). Each of these acid addition salts can be either amono-addition salt or a poly-addition salt, as these terms are definedherein.

Depending on the stoichiometric proportions between the charged group(s)in the compound and the counter-ion in the salt, the acid or baseadditions salts can be either mono-addition salts or poly-additionsalts.

The phrase “mono-addition salt”, as used herein, refers to a salt inwhich the stoichiometric ratio between the counter-ion and charged formof the compound is 1:1, such that the addition salt includes one molarequivalent of the counter-ion per one molar equivalent of the compound.

The phrase “poly-addition salt”, as used herein, refers to a salt inwhich the stoichiometric ratio between the counter-ion and the chargedform of the compound is greater than 1:1 and is, for example, 2:1, 3:1,4:1 and so on, such that the addition salt includes two or more molarequivalents of the counter-ion per one molar equivalent of the compound.

As used herein, the term “prodrug” refers to a compound which isconverted in the body to an active compound (e.g., the compound of theformula described hereinabove). A prodrug is typically designed tofacilitate administration, e.g., by enhancing absorption. A prodrug maycomprise, for example, the active compound modified with ester groups,for example, wherein any one or more of the hydroxyl groups of acompound is modified by an acyl group, optionally (C₁₋₄)acyl (e.g.,acetyl) group to form an ester group, and/or any one or more of thecarboxylic acid groups of the compound is modified by an alkoxy oraryloxy group, optionally (C₁₋₄)alkoxy (e.g., methyl, ethyl) group toform an ester group.

Further, each of the compounds described herein, including the saltsthereof, can be in a form of a solvate or a hydrate thereof.

The term “solvate” refers to a complex of variable stoichiometry (e.g.,di-, tri-, tetra-, penta-, hexa-, and so on), which is formed by asolute (the heterocyclic compounds described herein) and a solvent,whereby the solvent does not interfere with the biological activity ofthe solute.

The term “hydrate” refers to a solvate, as defined hereinabove, wherethe solvent is water.

The compounds described herein can be used as polymorphs and the presentembodiments further encompass any isomorph of the compounds and anycombination thereof.

The present embodiments further encompass any enantiomers anddiastereomers of the compounds described herein.

As used herein, the term “enantiomer” refers to a stereoisomer of acompound that is superposable with respect to its counterpart only by acomplete inversion/reflection (mirror image) of each other. Enantiomersare said to have “handedness” since they refer to each other like theright and left hand. Enantiomers have identical chemical and physicalproperties except when present in an environment which by itself hashandedness, such as all living systems. In the context of the presentembodiments, a compound may exhibit one or more chiral centers, each ofwhich exhibiting an R- or an S-configuration and any combination, andcompounds according to some embodiments of the present invention, canhave any their chiral centers exhibit an R- or an S-configuration.

The term “diastereomers”, as used herein, refers to stereoisomers thatare not enantiomers to one another. Diastereomerism occurs when two ormore stereoisomers of a compound have different configurations at one ormore, but not all of the equivalent (related) stereocenters and are notmirror images of each other. When two diastereoisomers differ from eachother at only one stereocenter they are epimers. Each stereo-center(chiral center) gives rise to two different configurations and thus totwo different stereoisomers. In the context of the present invention,embodiments of the present invention encompass compounds with multiplechiral centers that occur in any combination of stereo-configuration,namely any diastereomer.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination or as suitable in any other describedembodiment of the invention. Certain features described in the contextof various embodiments are not to be considered essential features ofthose embodiments, unless the embodiment is inoperative without thoseelements.

Various embodiments and aspects of the present invention as delineatedhereinabove and as claimed in the claims section below find experimentalsupport in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with theabove descriptions illustrate some embodiments of the invention in anon-limiting fashion.

MATERIALS AND METHODS

Materials:

Anti-human IgG-XL665 antibody was obtained from Cisbio Bioassays.

Ficoll Histopaque® 1077 was obtained from Sigma (Israel).

Biotin was obtained from Sigma (Israel).

Biotinylated MIP3a was obtained from Almac Sciences (UK).

Terbium cryptate-conjugated streptavidin (Lumi4®) was obtained fromCisbio Bioassays.

BKT130 was prepared as described in International Applicationpublication WO2010/146584, in which BKT130 is referred to as“BKT-P2-FC”. The sequence of BKT130 is also presented therein.

Compound BKT300 was obtained from AnalytiCon Discovery GmbH at 78%purity and at high purity. Using NMR spectroscopy, the high puritysample was determined to have a purity of about 98%, whereas the othersample was confirmed to have about 78% purity.

Migration Assay:

600 μl of RPMI medium was added to the lower chambers of Transwell®transmigration plates, supplemented with 2 μg/ml of MIP3a, 100 ng/ml ofSDF-1 or 10 ng/ml of MCP-1. The tested small molecule was added to thelower chambers at the indicated concentration, except in controlsamples. The MIP3a, SDF-1 or MCP-1 was incubated with the small moleculefor 30 minutes at room temperature before the initiation of themigration assay. Following 30 minutes of incubation 2×10⁵ immune cellswere added to the upper chambers of the transmigration plates in a totalvolume of 100 μl. Cells which migrated within 3 hours to the bottomchamber of the Transwell® plates were counted using a FACScalibur™ flowcytometer.

To evaluate migration toward MIP3a, peripheral blood mononuclear cells(PBMCs) were isolated from heparinized venous blood by centrifugationover Ficoll Histopaque® 1077. CD4+ T cells were further isolated withRosetteSep™ human CD4+ T-cell Enrichment cocktail (StemCell TechnologiesInc.), according to the manufacturer's instructions. CD4+ T cells werere-suspended in RPMI medium containing 1% fetal calf serum (FCS).

To evaluate migration towards SDF-1, Jurkat cells were re-suspended inRPMI medium containing 1% fetal calf serum (FCS).

To evaluate migration towards MCP-1, THP-1 cells were re-suspended inRPMI medium containing 1% fetal calf serum (FCS).

Example 1 Screening Assay and Activity Assays Identifying SmallMolecules which Bind to and Affect Migration of MIP3a

A homogeneous time-resolved fluorescence (HTRF) assay was designed as aplatform for high-throughput screening (HTS). This assay detected theinteraction of BKT130 with MIP3a, using BKT130, biotinylated MIP3a,anti-human IgG-XL665 antibody (which binds to the Fc domain of BKT130)and Lumi4® terbium cryptate-conjugated streptavidin (which binds to thebiotin moiety attached to MIP3a).

Biotinylated MIP3a or biotin was diluted in an assay buffer of phosphatebuffer saline (PBS) with 0.1% bovine serum albumin (BSA) to a finalconcentration of 16.7 nM. A detection mix was formed by diluting BKT130,terbium-conjugated streptavidin and anti-human IgG-XL665 antibody in theassay buffer to concentrations of 92.5 nM, 0.01 ng/ml, and 0.9 ng/ml,respectively. 23 μl reactions were incubated in black non-binding384-well plates (Greiner 784900) at room temperature for 45 minutes, andthen read in a PHERAstar FS® high-throughput microplate reader (BMGLABTECH) with a dedicated HTRF laser excitation. HTRF reads are afunction of resonant energy transfer from the terbium donor (emitting ata wavelength of 625) to the XL665 acceptor, which becomes excited andemits a fluorescent signal at a wavelength of 665 nm. Onlydonor/acceptor pairs that are brought into close proximity by binding ofMIP3a to BKT130 will result in resonant energy transfer. Binding isexpressed as the ratio of the signal High-throughput screening (HTS) wasperformed using an automated workstation with integrated 50 nl pin tooland BioTek™ EL406 dispenser. Compounds from a natural library of about3,500 compounds were maintained in DMSO stock solutions of approximately10 mM and then transferred to an assay mix containing biotin-MIP3a (orbiotin control), and incubated for 15 minutes at room temperature toallow for compound binding. The detection mix was then added; plateswere then incubated for a further 45 minutes at room temperature, andthen read as described hereinabove.

Compounds with a significant inhibition of binding were selected andpicked from the library for repeat assays in serial dilution to obtaindose response curves.

Analysis of the screen and curve fitting was done using the GenedataScreener® software package.

In the presence of BKT130 and biotinylated-MIP3a without any additionalcompounds, the signal ratio was 3621 (±409), which corresponded to 0%inhibition of binding (neutral control). In the presence of biotin aloneand BKT130, the obtained signal ratio was 763 (±23), corresponding to100% inhibition of binding (inhibitor control).

FIG. 1 shows the distribution of signals obtained from the HTS of the3,500 compounds.

As shown in FIG. 1 and in Table 1, 32 of the 3,500 screened compoundsinhibited the binding of BKT130 to MIP3a (as expressed by the ratio ofthe signal at 665 nm to the signal at 625 nm) by more than 40%.

TABLE 1 Effect of 32 most inhibitory compounds on binding of BKT130 toMIP3a Compound FRET Change in No. Signal Ratio binding 1 915 −92.2% 2946 −91.5% 3 1040 −86.3% 4 1265 −78.8% 5 1320 −77.1% 6 1528 −68.6% 71692 −60.2% 8 1615 −60.0% 9 1751 −59.7% 10 1722 −59.7% 11 1777 −58.1% 121748 −57.7% 13 1797 −57.5% 14 1829 −56.2% 15 1814 −54.9% 16 1721 −54.2%17 1842 −54.1% 18 1834 −53.9% 19 1927 −52.2% 20 1779 −51.5% 21 1911−51.2% 22 1911 −50.7% 23 1945 −49.8% 24 2004 −49.3% 25 1793 −49.3% 262040 −47.5% 27 2021 −46.6% 28 2033 −46.1% 29 2098 −45.4% 30 1918 −43.2%31 2143 −41.5% 32 2148 −41.5%

Of these 32 small molecules, 18 small molecules were found to bothsignificantly inhibit the interaction between BKT130 and MIP3a in thehigh-throughput screening and showed a dose response curve in the serialdilution assay, and were selected for further analysis. The structuresof the 18 small molecules are presented in Table 2 below.

TABLE 2 Compounds which exhibited dose response curve for MIP3a bindingComp. No. Name Structure BKT200

BKT201

BKT202

BKT203 Hypericin

BKT204 Cyrtominetin

BKT205

BKT206

BKT207 Aloesaponarin I

BKT208 Trichoclin

BKT209

BKT210 Sennoside A

BKT211

BKT212 Falcarindiol

BKT213

BKT214

BKT215 Auraptene

BKT216 Arjunolic acid

BKT300

The 18 compounds uncovered by the screening assay were further testedfor their ability, at final concentrations of 10 and 50 μg/ml, toinhibit the migration of human CD4+ T-cells toward MIP3a; using theprocedures described in the Materials and Methods section hereinabove.

The results for 3 exemplary compounds presented in Table 2, CompoundsBKT210, BKT203 and BKT207, which exhibited inhibition of CD4+ T-cellsmigration toward MIP3a, are presented herein.

FIGS. 2A-2C present the dose response curves of Compounds BKT210, BKT203and BKT207, respectively.

As shown in FIGS. 3-9, Compounds BKT300 (at 78% purity) (FIG. 3), BKT201(FIG. 4), BKT202 (FIG. 5), BKT205 (FIG. 6), BKT209 (FIG. 7), BKT210(FIG. 8) and BKT213 (FIG. 9), at a concentration of 50 μg/ml, completelyinhibited CD4+ T-cell migration towards MIP3a, thus indicating that thebinding of the compound to MIP3a (as detected in the HTS assay) isassociated with inhibition of MIP3a activity. As further shown in FIGS.5 and 8, Compounds BKT202 and BKT210, at a concentration of 10 μg/ml,significantly inhibited CD4+ T-cell migration towards MIP3a.

As shown in FIGS. 10-13, Compounds BKT203 (FIG. 10), BKT206 (FIG. 11),BKT207 (FIG. 12) and BKT212 (FIG. 13), at a concentration of 10 μg/ml,each completely inhibited CD4+ T-cell migration towards MIP3a.

These results confirm that the binding of the compounds to MIP3a (asdetected in the HTS assay) is associated with inhibition of MIP3aactivity.

Example 2 Effect of Exemplary Compounds on Cytokine-Induced Migration

The set of 18 small molecules presented in Table 2 was screened for anability to inhibit the migration of immune cells in response to MCP-1and SDF-1, using the procedures described in the Materials and Methodssection hereinabove, in accordance with the screening process describedin Example 1.

FIGS. 14A and 14B show that BKT300, at a purity of 78% (FIG. 14A) and of98% (FIG. 14B) at a concentration of 10 μg/ml, significantly inhibitedthe migration of lymphocytic Jurkat cells towards SDF-1.

A compound structurally similar to BKT300, termed BKT400, was also foundto inhibit the migration of lymphocytic Jurkat cells towards SDF-1, asshown in FIG. 15.

Furthermore, as shown in FIGS. 16-27, BKT201 (FIG. 16), BKT203 (FIG.17), BKT204 (FIG. 18), BKT205 (FIG. 19), BKT206 (FIG. 20), BKT207 (FIG.21), BKT208 (FIG. 22), BKT209 (FIG. 23), BKT210 (FIG. 24), BKT211 (FIG.25), BKT212 (FIG. 26) and BKT213 (FIG. 27), at a concentration of 10μg/ml, significantly inhibited the migration of lymphocytic Jurkat cellstowards SDF-1, with BKT201, BKT203, BKT205, BKT206, BKT207, BKT209,BKT211, BKT212 and BKT213 exhibiting strong inhibition at thisconcentration.

These results indicate that the abovementioned compounds are effectiveinhibitors of SDF-1 function, and suggest that these compounds areeffective for treating conditions associated with activity of SDF-1 andCXCR4 (the receptor of SDF-1).

Furthermore, as shown in FIGS. 28-35, Compounds BKT300 (78% purity)(FIG. 28), BKT201 (FIG. 29), BKT204 (FIG. 30), BKT205 (FIG. 31), BKT206(FIG. 32), BKT209 (FIG. 33), BKT211 (FIG. 34) and BKT216 (FIG. 35), at aconcentration of 50 μg/ml, exhibited strong inhibition of migration ofmonocytic THP-1 cells towards MCP-1.

As shown in FIGS. 28, 29, 31 and 33, at a concentration of 10 μg/ml,Compounds BKT300 (78% purity) (FIG. 28), BKT201 (FIG. 29), BKT205 (FIG.31) and BKT209 (FIG. 33) had no apparent effect on the migration ofmonocytic THP-1 cells towards MCP-1.

As shown in FIGS. 30, 32 and 35, at a concentration of 10 μg/ml,Compounds BKT204 (FIG. 30), BKT206 (FIG. 32) and BKT216 (FIG. 35)exhibited partial inhibition of migration towards MCP-1.

As shown in FIG. 34, 10 μg/ml of Compound BKT211 exhibited completeinhibition of THP-1 cell migration towards MCP-1. This result suggeststhat Compound BKT211 is effective for treating conditions associatedwith activity of MCP-1.

Taken together with the results described in Example 1 (e.g., as shownin FIGS. 3, 4, 6, 7 and 9), the above results indicate that CompoundsBKT300, BKT201, BKT205, BKT209 and BKT213 potently inhibit SDF-1function in a relatively selective manner, with considerably weakerinhibition of MCP-1 and/or MIP3a function (e.g., at a concentration ofabout 10 μg/ml), and suggest that Compounds BKT300, BKT201, BKT205,BKT209 and BKT213 are particularly effective for treating conditionsassociated with activity of SDF-1 and CXCR4 (the receptor of SDF-1).

Example 3 Effect of Exemplary Compounds on Cancer Cell Viability

In order to assess the effect of compounds of Table 2 on cancer cellviability, the in vitro effect of the compounds on MV4-11 human acutemyeloid leukemia cells was evaluated. The cancer cells were incubated inRPMI cell medium with 1% fetal calf serum (FCS) at a concentration of2×10⁵ cells/well at a final volume of 250 μl in 96-well plate. Thecompounds were added to the cells at the indicated concentrations. Thecells were incubated for 24 hours and the number of dead and viablecells was then evaluated by fluorescence-activated cell sorting (FACS),using propidium iodide (PI) staining. The IC50 of chemokine-induced celldeath was determined using GraphPad Prism software.

As shown in FIGS. 36A and 36B, Compound BKT206 induced cell death ofMV4-11 human acute myeloid leukemia cells at concentrations of 10 μg/ml.

As shown in FIGS. 37A and 37B, Compound BKT211 induced cell death ofMV4-11 human acute myeloid leukemia cells at concentrations of 10 μg/ml.

As shown in FIGS. 38A and 38B, Compound BKT215 induced cell death ofMV4-11 human acute myeloid leukemia cells at concentrations of 20 μm(equivalent to 6 μg/ml).

As shown in FIGS. 39A and 39B, BKT300 (at 78% purity) induced cell deathof MV4-11 human acute myeloid leukemia cells at concentrations as low as2 μg/ml (the lowest tested concentration). The IC₅₀ for BKT300 (78%purity)-induced death of MV4-11 cells was 2.72 μg/ml.

Induction of cell death by BKT300 was repeated while comparing theeffects of a sample of BKT300 at 78% purity with a sample of BKT300 at98% purity.

As shown in FIGS. 40A-40D, BKT300 at 98% purity was significantly moreeffective than BKT300 at 78% purity at inducing cell death. For example,BKT300 at 98% purity induced a considerably greater degree of cell deathat 4.25 μg/ml than did BKT300 at 78% purity.

These results suggest that impurities present in BKT300 (e.g., BKT300 at78% purity) may reduce the cell death induced by BKT300 per se, and thatBKT300 at a high degree of purity (e.g., 98%) is particularly potent incomparison to other compounds described herein.

These results indicate that chemokine-binding activity of compoundsdescribed herein is associated with anticancer activity.

The in vitro effect of BKT300 (at 78% purity) on cancer cell viabilitywas further evaluated using a variety of additional cancer cell lines,using procedures as described hereinabove with respect to MV4-11 cells.

As shown in FIGS. 41A and 41B, BKT300 (at 78% purity) induced cell deathof RPMI human multiple myeloma cells at concentrations as low as 2 μg/ml(the lowest tested concentration).

As shown in FIGS. 42A and 42B, BKT300 (at 78% purity) induced cell deathof Jurkat human acute lymphoblastic leukemia cells at concentrations aslow as 2 μg/ml (the lowest tested concentration). The IC50 forBKT300-induced death of Jurkat cells was 3.5 μg/ml.

As shown in FIGS. 43A-44B, BKT300 (at 78% purity) induced cell death ofabout 30% of Raji (FIGS. 43A and 43B) and Bjab (FIGS. 44A and 44B) humanlymphoma cells at a concentration of 8.5 μg/ml, and further inducedslight cell death of Raji cells at a concentration of 4.25 μg/ml (FIGS.43A and 43B).

As shown in FIGS. 45A and 45B, BKT300 (at 78% purity) induced cell deathof about 80% of H460 human large cell lung cancer cells at aconcentration of 10 μg/ml.

As shown in FIGS. 46A and 46B, BKT300 (at 78% purity) induced cell deathof over 90% of H345 human small cell lung cancer cells at aconcentration of 8.5 μg/ml.

These results indicate that BKT300 is effective at inducing cell deathof a wide variety of cancer cell types.

Example 4 Effect of Exemplary Compound on Cancer Cells In Vivo

The effect of BKT300 (at 98% purity) on the proliferation of AML cellsin vivo was examined by treatment of NOD scid gamma (NSG) micetransplanted with MV4-11 (FLT3-ITD) cells.

The mice were subjected to irradiation with 300 rad and on the followingday were transplanted by IV injection with MV4-11 (FLT3-ITD) cells,10×10⁶ cells/mouse. 21 days following transplantation, the treated groupwas injected intraperitoneally with 1 mg/Kg of BKT300 (98% purity) perinjection for three consecutive days. On day 25 followingtransplantation, mice were sacrificed and the survival of the human AMLblasts in the blood, spleen and the bone marrow was evaluated using antihuman CD45.

The study protocol is presented in the following table and some of theresults are presented in FIGS. 47A-47C.

Treatments Day Day Day Day End of Exp. Day (−1) Day 0 21 22 23 24 Day 25Irradiation Cells Mice sacrifice 300 rad transplantation Blood 10 × 10⁶BM MV4-11 (IV) 1 + + + + spleen mg/Kg/mouse

As shown in FIG. 47B, BKT300 administration dramatically reduced thenumber and percentage of AML cells in the bone marrow of mice.

FIGS. 47B and 47C present data obtained in a representative FACSanalysis showing the presence of human MV4-11 cells with in the bonemarrow of untreated mice (FIG. 47C) and of mice treated with BKT300,which further demonstrate the ability of BKT300 to eradicate theleukemic cells.

Example 5 Inhibition of Kinase Activity by BKT300

In order to further characterize the effect of BKT300 on cell signaling,kinase profiling of BKT300 (at 78% purity) was performed (by the LifeTechnologies SelectScreen® Biochemical Profiling Lab) using aLanthaScreen® europium kinase binding assay to screen 440 kinases.

The principle of the LanthaScreen® assay is depicted in FIG. 48. Bindingof an Alexa Fluor® conjugate or “tracer” to a kinase is detected byaddition of a europium (Eu)-labeled anti-tag antibody. Binding of thetracer and antibody to a kinase results in a high degree of FRET,whereas displacement of the tracer with a kinase inhibitor results in aloss of FRET. The kinase tracers are based on ATP-competitive kinaseinhibitors, making them suitable for detection of any compounds thatbind to the ATP site. Inhibitors that bind the ATP site include bothType I kinase inhibitors, which bind solely to the ATP site, and Type IIinhibitors (e.g., imatinib, sorafenib, BIRB-796), which bind to both theATP site and a second site often referred to as the allosteric site.

Of the 440 screened kinases, BKT300 inhibited 36 kinases by more than40%. These kinases are presented in Table 3 below.

As shown in Table 3, most of the kinases inhibited by BKT300 wereserine/threonine kinases.

Many such kinases are involved in cancer, and some in immune regulation.These results suggest that kinase inhibition by BKT300 can be utilizedfor treating cancer, particularly by cancer immunotherapy.

TABLE 3 Inhibition of kinases by BKT300 % Kinase Kinase Inhibition TypeDYRK3 47 ST EPHA8 50 ND GRK4 63 ST GRK5 65 ST MAP4K2 (GCK) 48 ND MAP4K4(HGK) 40 ST MELK 41 ST PAK7 (KIAA1264) 40 ST SGK2 43 ST SRC N1 41 TKACVRL1 (ALK1) 47 ST BMPR1A (ALK3) 58 ST CDC7/DBF4 53 ST CDK1/cyclin A245 ST CDK11 (Inactive) 57 ST CDK8/cyclin C 64 ND CLK4 73 ST DAPK2 65 STDYRK2 62 ST ICK 41 ST KIT D820E 42 TK* KIT T670E 51 TK* MAP4K1 (HPK1) 45ST MAPK10 (JNK3) 49 ST MLCK (MLCK2) 58 ST MYLK (MLCK) 63 ST NUAK2 89 STSTK17A (DRAK1) 48 ST STK17B (DRAK2) 107 ST STK38 (NDR) 41 ST STK38L(NDR2) 45 ST TGFBR2 43 ST TTK 52 STTK DAPK1 43 ST PIK3CA 64 PIK3CD 77 ST= serine/threonine kinase TK = tyrosine kinase STTK = serine/threoninetyrosine kinase ND = kinase type not determined

Example 6 Computational Binding Model of BKT300 to Kinases

All modeling work was performed using the Accelrys software package“Discovery Studio”.

Pharmacophore models were constructed manually (not using the automatedpharmacophore tools of the package).

All small molecule conformations were generated using the “BEST”conformational search algorithm.

Pharmacophore mapping was performed using the “Pharmacophore mapping”tool of Discovery Studio, with the “flexible” option turned on.

All results of pharmacophore mapping were visually inspected in order tochoose best candidate poses.

Design of a Binding Model to Kinases:

As demonstrated in Examples 1-4 hereinabove, BKT300 was identifiedthrough a cell-based assay as a promising active agent against leukemiacell lines. As shown in Example 5 hereinabove, in a screen of inhibitionagainst the human kinome it was shown to inhibit a selection of kinases.Based on this inhibition data, coupled with gene expression data andbiological considerations, four kinases were chosen as potential targetsthat could, possibly in some combination, mediate the anti-leukemiaeffect of BKT300: MELK, MAP4K4 and two Pi3-kinases (Pik3Cα and Pik3Cδ;also referred to as PIK3CA and PIK3CD), highlighted in Table 3hereinabove.

Structural analysis of these four kinases was performed using allavailable structures in the public domain (PDB). For a preliminaryconstruction of pharmacophoric models, the two protein-kinases, MELK andMAP4K4, were selected.

A literature search was performed to identify experimentally-verified“hot spots” (amino acid residues that if mutated result in loss of anorder of magnitude or more in activity) for each of the kinases. Twosuch amino acid residues were identified: Lys40 and Asp150, bothpositioned within the ATP binding site of the kinases.

The two protein kinases were then aligned so as to achieve the bestpossible alignment of the ATP binding pocket, and in particular of Lys40and Asp150. The alignment is shown in FIG. 49.

Inhibitors of these two kinases known in the art were used both todevelop a binding model, and to develop a scoring function for rankingpotential compounds with respect to their predicted ability to inhibitMELK and MAP4K4.

Two datasets were compiled: (i) a dataset of MELK inhibitors whichincludes 76 compounds with affinities to the enzyme in the range of from4.9 to more than 10000 nM; and (ii) a dataset of MAPK4K inhibitors whichincludes 8 compounds with affinities to the enzyme in the range of from140 to more than 10000 nM.

Using the crystal structures of available MELK and MAPK4K inhibitors, abinding model that contains a pharmacophore, and overall shape of theligands were constructed. The pharmacophore was designed such that thebound ligands are required to interact with Lys40 and Asp150.

Validation of the model was performed by mapping the known MELK andMAPK4K inhibitors from the above datasets onto the model. 90% of all ofthe evaluated MELK inhibitors were successfully mapped to thepharmacophore, whereby for the high affinity inhibitors, featuring KDlower than 1000 nM, 100% were successfully mapped onto the model. Forthe MAPK4K inhibitors, all of the 8 inhibitors were successfully mappedto the model.

These results indicated that the designed binding model is valid and canbe used in predicting the binding mode of BKT300.

Predicting the Binding Conformation of BKT300 to Kinases:

BKT300 was mapped to the designed binding model: all low energyconformations of BKT300 were generated, and mapped to the model. Allsuccessfully mapped conformers were then docked to the binding site ofMELK using the model as a guide, and the docked complex wasenergetically minimized, allowing the side chains of the protein toadjust to each pose. 165 successful conformations/poses were obtained,and each was visually inspected to evaluate the interaction of theligand with MELK, to thereby select the most suitable conformation andpose, which is depicted in FIG. 50.

In order to provide additional support for this pose, known crystalstructures of MELK inhibitors were screened in order to identifycompounds that feature groups that occupy the same positions of thekinase as do the aliphatic groups (“tails”) of BKT300, flanking the3-ring skeleton.

Two such structures were found:N-[3-(4-aminoquinazolin-6-yl)-5-fluorophenyl]-2-(pyrrolidin-1-yl)acetamide(PDB 40BQ) and3′-{[(4-bromo-1-methyl-1H-pyrrol-2-yl)carbonyl]amino}-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]-1′,4′-dihydro-5′H-spiro[cyclopropane-1,6′-pyrrolo[3,4-c]pyrazole]-5′-carboxamide(PDB 4BKY). These structures were overlaid on the selected pose ofBKT300, as shown in FIG. 51.

It is noted that the chemical nature of the flanking groups (“tails”) ofthese inhibitors differ from the flanking alkyl groups of BKT300, yetoccupy the same sub-pockets in the protein kinase. It is further notedthat the affinity of BKT300 is relatively low (few tens of μM based onthe kinase screening assay summarized in Table 3 hereinabove), wherebythe affinities of the overlaid inhibitors is significantly higher (atthe nM range).

Example 7 Preparation of BKT300 Analogs

Using the above-described binding model, catalogs of more than 40million compounds were screened in order to identify compounds thatfeature the same topology (same spatial arrangement/conformation) as theselected pose of BKT300 shown in FIG. 51. Each compound that matched theBKT300 topology was mapped onto the binding model, and its ability tofit the MELK and MAPK4K binding site was evaluated by visual inspection.The top scoring compounds constituted a set of about 20 compounds, thestructures of which are presented in Table 4 below.

Notably, all of the retrieved compounds were of the same supplier,Angene Chemical, yet were found to be commercially unavailable.

TABLE 4 PubChem Compund ID PUBCHEM_IUPAC_CAS_NAME Structure 235640034-[(4-fluoro-6-methyl-1,3,5,-triazin- 2-yl)amino]-5-hydroxy-7-(trihydroxy-$l{circumflex over ( )}{4}-sulfanyl)-2- naphthalenesulfonicacid

25226044 2,4-diamino-5,7-dianilino-8-chloro- 6-quinazolinecarbonitrile

59055724 4-[[4-chloro-6-(1- naphthalenylamino)-1,3,5-triazin-2-yl]amino]-5-hydroxy-7-(trihydroxy- $l{circumflex over( )}{4}-sulfanyl)-1- naphthalenesulfonic acid

59055720 4-[[4-chloro-6-(1- naphthalenylamino)-1,3,5-triazin-2-yl]amino]-5-hydroxy-7-(trihydroxy- $l{circumflex over( )}{4}-sulfanyl)-2- naphthalenesulfonic acid

59372660 5-[(4,6-dimethyl-1,3,5-triazin-2- yl)amino]-4-hydroxy-2-naphthalenesulfenic acid hydroperoxy ester

15480413 (5-benzoyl-4,8-dihydroxy-1- naphthalenyl)-phenylmethanone

19771237 4-[[4-(2-aminoethylamino)-6-chloro-1,3,5-triazin-2-yl]amino]-5- hydroxynaphthalene-2,7-disulfonicacid

25226217 2,4-diamino-8-chloro-5,7- bis(phenylthio)-6-quinazolinecarbonitrile

23038808 acetic acid [8-(2,6- dichlorophenyl)sulfinyl-4-hydroxy-2,3-dimethoxy-1-naphthalenyl] ester

49862178 2,4-diamino-8-chloro-5,7- diphenoxy-6-quinazolinecarbonitrile

25226218 2,4-diamino-8-fluoro-5,7- bis(phenylthio)-6-quinazolinecarbonitrile

23038659 acetic acid [8-(2- bromophenyl)sulfinyl-4-hydroxy-2,3-dimethoxy-1-naphthalenyl] ester

19090340 5-[[4-fluoro-6-(2-sulfoethylamino)-1,3,5-triazin-2-yl]amino]-4-hydroxy- 2-naphthalenesulfonic acid

11245388 8-[(4-hydroxy-1-naphthalenyl)oxy]-4-(methoxymethoxy)-1-naphthalenol

17891057 5-[(2,4-diamino-5- pyrimidinyl)methyl]-2,3-dimethoxy-4-(2-phenylethynyl)naphthalene-1,7- diol

17903633 4-(1H-indazol-5-ylamino)-5,7- dimethoxy-3-quinolinecarbonitrile

53738023 2-[4-hydroxy-3-methoxy-5- (phenylmethyl)-1-naphthalenyl]-3-phenyl-2-propenoic acid

21149542 4-[(2-aminophenyl)-oxomethyl]-5-hydroxynaphthalene-1,7-disulfonic acid

1895649 (5-benzoyl-4-hydroxy-8- methoxynaphthalen-1-yl)- phenylmethanone

In parallel, compounds which are structural analogs of BKT300 weredesigned, the structures of which are presented in Table 5 below.

TABLE 5 Molecule Name

BKT300-1

BKT300-2

BKT300-3

BKT300-4

BKT300-5

BKT300-6

BKT300-7

BKT300-8

BKT300-9

BKT300-10

BKT300-11

BKT300-12

BKT300-13

BKT300-14

BKT300-15

BKT300-16

BKT300-17

BKT300-18

BKT300-19

BKT300-20

BKT300-21

BKT300-22

BKT300-23

Compounds BKT300-7, BKT300-23, BKT300-1, BKT300-3 and BKT300-11 (Table5) were synthesized as depicted in FIGS. 52A-B, 53A-B and 54-56,respectively. In addition, the preparation of the exemplary CompoundsBKT300-1, BKT300-3 and BKT300-11 is described in detail herein below.

The compounds' structures were verified by LC-MS and NMR.

Other compounds can be similarly prepared by using correspondingreactants.

Preparation of BKT300-1:

A scheme presenting the synthesis of BKT300-1 is presented in FIG. 54.

1. Preparation of1,5-dimethoxy-2-(5-methoxy-2-nitrophenoxy)-3-pentylbenzene (BKT300-1-b4)

To a solution of 2,4-dimethoxy-6-pentylphenol (b3) (2.2 grams, 0.981mmol) in tetrahydrofuran (THF) (20 ml) was added NaH (60%) (78.48 mg,1.962 mmol). The reaction mixture was stirred at 0° C. for 30 minutes.Then, 2-fluoro-4-methoxy-1-nitrobenzene (1.68 gram, 0.981 mmol) wasadded at 0° C. The reaction mixture was stirred at room temperatureovernight. TLC (thin layer chromatography) showed the reaction wascompleted (EtOAc: Petroleum Ether=1:10). The reaction mixture was pouredinto ice-water and extracted with EtOAc (2×20 ml). The organic layer waswashed with brine (2×50 ml), dried over anhydrous Na₂SO₄, and filtered.The filtrate was concentrated in vacuum to give the product1,5-dimethoxy-2-(5-methoxy-2-nitrophenoxy)-3-pentylbenzene (BKT300-1-b4)as a yellow oil (1.86 gram, 50.5% yield).

2. Preparation of 2-(2,4-dimethoxy-6-pentylphenoxy)-4-methoxyaniline(BKT300-1-b5)

A mixture of 1,5-dimethoxy-2-(5-methoxy-2-nitrophenoxy)-3-pentyl benzene(BKT300-1-b4) (1.86 gram, 4.95 mmol) and Raney nickel (200 mg) in MeOH(20 ml) was stirred at room temperature for 4 hours. LC-MS (liquidchromatography-mass spectroscopy) showed the reaction was completed. Thereaction mixture was filtered. The filtrate was concentrated in vacuumto give the product 2-(2,4-dimethoxy-6-pentylphen oxy)-4-methoxyaniline(BKT300-1-b5) as a black oil (1.21 gram, 70.7% yield). LC-MS: m/z 346.0(M++H).

3. Preparation of methyl2-((2-(2,4-dimethoxy-6-pentylphenoxy)-5-methoxyphenyl)carbamoyl)heptanoate(BKT300-1-b6)

A mixture of 2-(2,4-dimethoxy-6-pentylphen oxy)-4-methoxyaniline(BKT300-1-b5) (1.21 gram, 3.50 mmol), dimethyl-2-pentylmalonate (1.416gram, 7.00 mmol) and pyridine (0.553 gram, 7.00 mmol) in toluene (20 ml)was stirred at reflux for 40 hours. LC-MS (liquid chromatography-massspectroscopy) showed the reaction was completed. The reaction mixturewas concentrated in vacuum. The residue was purified by silica gelchromatography eluted with (EtOAc: Petroleum Ether-1:201:10) to give theproduct methyl 2-((2-(2,4-dimethoxy-6-pentylphenoxy)-5-methoxyphenyl)carbamoyl)heptanoate (BKT300-1-b6) as a yellowoil (1.882 gram, 100% yield). LC-MS: m/z 516.0 (M++H).

4. Preparation of2-((2-(2,4-dimethoxy-6-pentylphenoxy)-5-methoxyphenyl)carbamoyl)heptanoicAcid (BKT300-1-b7)

To a solution of methyl 2-((2-(2,4-dimethoxy-6-pentylphenoxy)-5-methoxyphenyl)carbamoyl)heptanoate (BKT300-1-b6) (1.882 gram,3.65 mmol) in a mixture solution of THF (10 ml), MeOH (10 ml) and water(10 ml) was added LiOH—H₂O (307 mg, 7.30 mmol). The reaction was stirredat room temperature for 16 hours. LC-MS (liquid chromatography-massspectroscopy) showed the reaction was completed. The reaction mixturewas concentrated in vacuum. The residue was dissolved in water (50 ml)and acidified to pH 2-3 using concentrated HCl. The reaction mixture wasextracted with EtOAc (2×20 ml). The organic layer was washed with brine(2×50 ml), dried over anhydrous Na₂SO₄, and filtered. The filtrate wasconcentrated in vacuum to give the product 2-((2-(2,4-dimethoxy-6-pentylphenoxy)-5-methoxyphenyl) carbamoyl)heptanoic acid (BKT300-1-b7) as ayellow solid (1.68 gram, 91.8% yield). LC-MS: m/z 502.0 (M++H).

5. Preparation of8-(2,4-dimethoxy-6-pentylphenoxy)-6-methoxy-3-pentylquinoline-2,4(1H,3H)-dione(BKT300-1-b8)

To a PPA solution (10 ml) at 120° C. was added2-((2-(2,4-dimethoxy-6-pentyl phenoxy)-5-methoxyphenyl)carbamoyl)heptanoic acid (BKT300-1-b7) (3.63 grams, 7.24 mmol)portion-wise. The reaction mixture was stirred at 120° C. for 2 hours.LC-MS (liquid chromatography-mass spectroscopy) showed the reaction wascompleted. The reaction mixture was poured into water (100 ml) andextracted with EtOAc (2×20 ml). The organic layer was washed with brine(2×50 ml), dried over anhydrous Na₂SO₄, and filtered. The filtrate wasconcentrated in vacuum. The residue was purified by silica gelchromatography eluted with (EtOAc:Petroleum Ether=1:101:5) to give theproduct 8-(2,4-dimethoxy-6-pentylphenoxy)-6-methoxy-3-pentylquinoline-2,4(1H,3H)-dione (BKT300-1-b8) as a yellow solid (320 mg, 8.67%yield). LC-MS: m/z 484.7 (M++H).

6. Preparation of2,4-dichloro-6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-1-b9)

A mixture of 8-(2,4-dimethoxy-6-pentylphenoxy)-6-methoxy-3-pentyl quinoline-2,4(1H,3H)-dione (BKT300-1-b8) (280 mg, 0.58 mmol) in POCl₃ (10 ml)was stirred at 80° C. for 6 hours. LC-MS (liquid chromatography-massspectroscopy) showed the reaction was completed. The reaction mixturewas concentrated in vacuum.

The residue was dissolved in EtOAc (20 ml) and washed successively withsaturated NaHCO₃(2×20 ml) and brine (20 ml), dried over anhydrousNa₂SO₄, and filtered. The filtrate was concentrated in vacuum to givethe product2,4-dichloro-6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-1-b9) as a light yellow solid (301 mg, 100% yield). LC-MS: m/z521.0 (M++H).

7. Preparation of4-chloro-8-(2,4-dimethoxy-6-pentylphenoxy)-2,6-dimethoxy-3-pentylquinoline(BKT300-1-b10)

A mixture of2,4-dichloro-6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-1-b9) (301 mg, 0.58 mmol) and MeONa (307 mg, 5.8 mmol) intoluene (10 ml) was stirred at 145° C. in a sealed tube for 21 hours.TLC showed the reaction was completed (EtOAc:Petroleum Ether=1:50). Thereaction mixture was concentrated in vacuum. Water was poured into theresidue and extracted with EtOAc (2×20 ml). The organic layer was washedwith brine (2×20 ml), dried over anhydrous Na₂SO₄, and filtered. Thefiltrate was concentrated in vacuum. The residue was purified by TLCeluted with (EtOAc:Petroleum Ether=1:100) to give the product4-chloro-8-(2,4-dimethoxy-6-pentylphenoxy)-2,6-dimethoxy-3-pentylquinoline(BKT300-1-b10) as a white solid (310 mg, 100% yield). LC-MS: m/z 516.0(M++H).

8. Preparation of8-(2,4-dimethoxy-6-pentylphenoxy)-2,6-dimethoxy-3-pentylquinoline(BKT300-1-b11)

A mixture of4-chloro-8-(2,4-dimethoxy-6-pentylphenoxy)-2,6-dimethoxy-3-pentylquinoline(BKT300-1-b10) (310 mg, 0.6 mmol) and 10% Pd/C (30 mg) in MeOH (20 ml)and Et₃N (2 ml) was stirred at room temperature for 2.0 hours. TLCshowed the reaction was completed (EtOAc:Petroleum Ether=1:20). Thereaction mixture was filtered. The filtrate was concentrated in vacuum.The residue was dissolved in EtOAc (20 ml) and washed with brine (2×20ml). The organic layer was dried over anhydrous Na₂SO₄, and filtered.The filtrate was concentrated in vacuum to give the product8-(2,4-dimethoxy-6-pentylphenoxy)-2,6-dimethoxy-3-pentyl quinoline(BKT300-1-b11) as a light yellow solid (300 mg, 100% yield). LC-MS: m/z482.1 (M++H).

9. Preparation of8-(2,4-dihydroxy-6-pentylphenoxy)-6-hydroxy-3-pentylquinolin-2(JH)-one(BKT300-1)

A mixture of 8-(2,4-dimethoxy-6-pentylphenoxy)-2,6-dimethoxy-3-pentylquinoline (BKT300-1-b11) (300 mg, 0.623 mmol) in 30% HBr in CH₃COOH (4mL) was stirred at 120° C. for 16 hours. Then the reaction mixture wasconcentrated in vacuum. The residue was dissolved in 30% HBr in CH₃COOH(4 ml) in a sealed tube and was stirred at 120° C. for 4 hours. Thereaction mixture was concentrated in vacuum. The residue was purified bypre-HPLC to give 8-(2,4-dihydroxy-6-pentylphenoxy)-6-hydroxy-3-pentylquinolin-2(1H)-one (BKT300-1) as a yellowsolid (77 mg, yield 29%). LC-MS: m/z 426.0 (M++H).

Preparation of BKT300-3:

A scheme presenting the synthesis of BKT300-3 is presented in FIG. 55.

1. Preparation of 2-(2,4-dimethoxyphenoxy)-4-methoxy-1-nitrobenzene(BKT300-3-c1)

To a solution of 2,4-dimethoxyphenol (R₁₁) (2.0 grams, 13.00 mmol) intetrahydrofuran (THF) (50 ml) was added NaH (60%) (450 mg, 26.00 mmol).The reaction mixture was stirred at 0° C. for 30 minutes. Then,2-fluoro-4-methoxy-1-nitrobenzene (2.22 grams, 13.00 mmol) was added at0° C. The reaction mixture was stirred at room temperature overnight.TLC showed the reaction was completed (EtOAc:Petroleum Ether=1:10). Thereaction mixture was poured into ice-water and extracted with EtOAc(2×20 ml). The organic layer was washed with brine (2×50 ml), dried overanhydrous Na₂SO₄, and filtered. The filtrate was concentrated in vacuumto give the product 2-(2,4-dimethoxyphenoxy)-4-methoxy-1-nitrobenzene(BKT300-3-c1) as a yellow oil (3.04 grams, 76.8% yield).

2. Preparation of 2-(2,4-dimethoxyphenoxy)-4-methoxyaniline(BKT300-3-c2)

A mixture of 2-(2,4-dimethoxyphenoxy)-4-methoxy-1-nitrobenzene(BKT300-3-c1) (3.04 grams, 10.00 mmol) and Raney nickel (770 mg) in MeOH(100 ml) was stirred at room temperature for 4 hours. LC-MS (liquidchromatography-mass spectroscopy) showed the reaction was completed. Thereaction mixture was filtered. The filtrate was concentrated in vacuumto give the product 2-(2,4-dimethoxyphenoxy)-4-methoxyaniline(BKT300-3-c2) as a black oil (2.58 grams, 94.2% yield). LC-MS: m/z 276.0(M++H).

3. Preparation of methyl 2-((2-(2,5-dimethoxyphenoxy)-5-methoxyphenyl)carbamoyl)heptanoate (BKT300-3-c3)

A mixture of 2-(2,4-dimethoxyphenoxy)-4-methoxyaniline (BKT300-3-c2)(3.75 grams, 13.62 mmol), dimethyl 2-pentylmalonate (5.5 grams, 27.2mmol) and pyridine (2.15 grams, 27.2 mmol) in toluene (40 ml) wasstirred at reflux for 40 hours. LC-MS (liquid chromatography-massspectroscopy) showed the reaction was completed. The reaction mixturewas concentrated in vacuum. The residue was purified by silica gelchromatography eluted with (EtOAc:Petroleum Ether-1:20-1:10) to give theproduct methyl 2-((2-(2,5-dimethoxyphenoxy)-5-methoxyphenyl)carbamoyl)heptanoate (BKT300-3-c3) as a yellow oil (5.0 grams, 82.45%yield). LC-MS: m/z 446.0 (M++H).

4. Preparation of2-((2-(2,4-dimethoxyphenoxy)-5-methoxyphenyl)carbamoyl)heptanoic Acid(BKT300-3-c4)

To a solution of methyl 2-((2-(2,5-dimethoxyphenoxy)-5-methoxyphenyl)carbamoyl)heptanoate (BKT300-3-c3) (5.0 grams, 11.23 mmol) in a mixturesolution of THF (10 ml), MeOH (10 ml) and water (10 ml) was addedLiOH—H₂O (944 mg, 22.46 mmol). The reaction was stirred at roomtemperature for 16 hours. LC-MS (liquid chromatography-massspectroscopy) showed the reaction was completed. The reaction mixturewas concentrated in vacuum. The residue was dissolved in water (50 ml)and acidified to pH 2-3 using concentrated HCl. The reaction mixture wasextracted with EtOAc (2×20 ml). The organic layer was washed with brine(2×50 ml), dried over anhydrous Na₂SO₄, and filtered. The filtrate wasconcentrated in vacuum to give the product2-((2-(2,4-dimethoxyphenoxy)-5-methoxyphenyl)carbamoyl)heptanoic acid(BKT300-3-c4) as a yellow solid (4.85 grams, 100% yield). LC-MS: m/z432.0 (M++H).

5. Preparation of8-(2,4-dimethoxyphenoxy)-6-methoxy-3-pentylquinoline-2,4(1H,3H)-dione(BKT300-3-c5)

To a PPA solution (8 mL) at 120° C. was added2-((2-(2,4-dimethoxyphenoxy)-5-methoxyphenyl)carbamoyl)heptanoic acid(BKT300-3-c4) (2.0 grams, 4.64 mmol) portion-wise. The reaction mixturewas stirred at 120° C. for 6 hours. LC-MS (liquid chromatography-massspectroscopy) showed the reaction was completed. The reaction mixturewas poured into water (100 ml) and extracted with EtOAc (2×20 ml). Theorganic layer was washed with brine (2×50 ml), dried over anhydrousNa₂SO₄, and filtered. The filtrate was concentrated in vacuum. Theresidue was purified by silica gel chromatography eluted with(EtOAc:Petroleum Ether=1:101:5) to give the product8-(2,4-dimethoxyphenoxy)-6-methoxy-3-pentylquinoline-2,4(1H,3H)-dione(BKT300-3-c5) as a yellow solid (240 mg, 12.5% yield). LC-MS: m/z 414.7(M++H).

6. Preparation of2,4-dichloro-8-(2,4-dimethoxyphenoxy)-6-methoxy-3-pentylquinoline(BKT300-3-c6)

A mixture of8-(2,4-dimethoxyphenoxy)-6-methoxy-3-pentylquinoline-2,4(1H,3H)-dione(BKT300-3-c5) (230 mg, 0.557 mmol) in POCl₃ (2 ml) was stirred at 110°C. for 4 hours. LC-MS (liquid chromatography-mass spectroscopy) showedthe reaction was completed. The reaction mixture was concentrated invacuum. The residue was dissolved in EtOAc (20 ml) and washedsuccessively with saturated NaHCO₃(2×20 ml) and brine (20 ml), driedover anhydrous Na₂SO₄, and filtered. The filtrate was concentrated invacuum to give the product 2,4-dichloro-8-(2,4-dimethoxyphenoxy)-6-methoxy-3-pentylquinoline (BKT300-3-c6) as a light yellowsolid (250 mg, 100% yield). LC-MS: m/z 451.0 (M++H).

7. Preparation of4-chloro-8-(2,4-dimethoxyphenoxy)-2,6-dimethoxy-3-pentylquinoline(BKT300-3-c7)

A mixture of 2,4-dichloro-8-(2,4-dimethoxy phenoxy)-6-methoxy-3-pentylquinoline (BKT300-3-c6) (250 mg, 0.55 mmol) and MeONa (300 mg, 5.5 mmol)in toluene (4 ml) was stirred at 145° C. in a sealed tube for 20 hours.

TLC showed the reaction was completed (EtOAc:Petroleum Ether=1:50). Thereaction mixture was concentrated in vacuum. Water was poured into theresidue and extracted with EtOAc (2×20 ml). The organic layer was washedwith brine (2×20 ml), dried over anhydrous Na₂SO₄, and filtered. Thefiltrate was concentrated in vacuum. The residue was purified by TLCeluted with (EtOAc:Petroleum Ether-1:100) to give the product4-chloro-8-(2,4-dimethoxyphenoxy)-2,6-dime thoxy-3-pentylquinoline(BKT300-3-c7) as a white solid (180 mg, 72.8% yield). LC-MS: m/z 447.0(M++H).

8. Preparation of8-(2,4-dimethoxyphenoxy)-2,6-dimethoxy-3-pentylquinoline (BKT300-3-c8)

A mixture of 4-chloro-8-(2,4-dimethoxyphenoxy)-2,6-dimethoxy-3-pentylquinoline (BKT300-3-c7) (180 mg, 0.40 mmol) and 10% Pd/C (20 mg) in MeOH(20 ml) and Et₃N (2 ml) was stirred at room temperature for 2.0 hours.TLC showed the reaction was completed (EtOAc:Petroleum Ether=1:20). Thereaction mixture was filtered. The filtrate was concentrated in vacuum.The residue was dissolved in EtOAc (20 ml) and washed with brine (2×20ml). The organic layer was dried over anhydrous Na₂SO₄, and filtered.The filtrate was concentrated in vacuum to give the product8-(2,4-dimethoxyphenoxy)-2,6-dimethoxy-3-pentylquinoline (BKT300-3-c8)as a light yellow solid (120 mg, 72.3% yield). LC-MS: m/z 412.1 (M++H).

9. Preparation of8-(2,4-dihydroxyphenoxy)-6-hydroxy-3-pentylquinolin-2(JH)-one (BKT300-3)

A mixture of 8-(2,4-dimethoxyphenoxy)-2,6-dimethoxy-3-pentylquinoline(BKT300-3-c8) (30 mg, 0.073 mmol) in 30% HBr in CH₃COOH (4 ml) wasstirred at 120° C. for 36 hours. The reaction mixture was thenconcentrated in vacuum. The residue and 10% Pd/C (2 mg) in MeOH (6 ml)was stirred at room temperature for 1 hour. The reaction mixture wasfiltered and the filtrate was concentrated in vacuum.

The residue was purified by pre-HPLC to give8-(2,4-dihydroxyphenoxy)-6-hydroxy-3-pentylquinolin-2(1H)-one (BKT300-3)as a yellow solid (4 mg, yield 15.3%). LC-MS: m/z 356.8 (M++H).

Preparation of BKT300-11:

A scheme presenting the synthesis of BKT300-11 is presented in FIG. 56.

1. Preparation of 4-methoxy-1-(5-methoxy-2-nitrophenoxy)-2-pentylbenzene(BKT300-11-a1)

To a solution of 4-methoxy-2-pentylphenol (R₁₁) (2.5 grams, 11.15 mmol)in THF (50 ml) was added NaH (60%) (892 mg, 22.30 mmol). The reactionmixture was stirred at 0° C. for 30 minutes. Then,2-fluoro-4-methoxy-1-nitrobenzene (1.91 gram, 11.15 mmol) was added at0° C. The reaction mixture was stirred at room temperature overnight.TLC showed the reaction was completed (EtOAc:Petroleum Ether=1:10). Thereaction mixture was poured into ice-water and extracted with EtOAc(2×20 ml). The organic layer was washed with brine (2×50 ml), dried overanhydrous Na₂SO₄, and filtered. The filtrate was concentrated in vacuumto give the product4-methoxy-1-(5-methoxy-2-nitrophenoxy)-2-pentylbenzene (BKT300-11-a1) asa yellow oil (3.85 grams, 100% yield).

2. Preparation of 4-methoxy-2-(4-methoxy-2-pentylphenoxy)aniline(BKT300-11-a2)

A mixture of 4-methoxy-1-(5-methoxy-2-nitrophenoxy)-2-pentylbenzene(BKT300-11-a1) (3.85 grams, 11.15 mmol, 1.0 eq.) and Raney nickel (770mg) in MeOH (100 ml) was stirred at room temperature for 4 hours. LC-MS(liquid chromatography-mass spectroscopy) showed the reaction wascompleted. The reaction mixture was filtered. The filtrate wasconcentrated in vacuum to give the product4-methoxy-2-(4-methoxy-2-pentyl phenoxy)aniline (BKT300-11-a2) as ablack oil (4.41 grams, 100% yield). LC-MS: m/z 316.0 (M++H).

3. Preparation of methyl 2-((5-methoxy-2-(5-methoxy-2-pentylphenoxy)phenyl)carbamoyl)heptanoate (BKT300-11-a3)

A mixture of 4-methoxy-1-(5-methoxy-2-nitrophenoxy)-2-pentylbenzene(BKT300-11-a1) (4.145 grams, 13.14 mmol), dimethyl 2-pentylmalonate(3.98 grams, 19.71 mmol) and pyridine (2.08 grams, 26.28 mmol) intoluene (80 ml) was stirred at reflux for 40 hours. LC-MS (liquidchromatography-mass spectroscopy) showed the reaction was completed. Thereaction mixture was concentrated in vacuum. The residue was purified bysilica gel chromatography eluted with (EtOAc:Petroleum Ether=1:20-1:10)to give the product methyl2-((5-methoxy-2-(5-methoxy-2-pentylphenoxy)phenyl)carbamoyl)heptanoate(BKT300-11-a3) as a yellow oil (7.2 grams, 100% yield). LC-MS: m/z 486.0(M++H).

4. Preparation of2-((5-methoxy-2-(4-methoxy-2-pentylphenoxy)phenyl)carbamoyl)heptanoicAcid (BKT300-11-a4)

To a solution of methyl 2-((5-methoxy-2-(5-methoxy-2-pentylphenoxy)phenyl)carbamoyl)heptanoate (BKT300-11-a3) (3.0 grams, 6.19mmol) in a mixture solution of THF (10 ml), MeOH (10 ml) and water (10ml) was added LiOH—H₂O (520 mg, 12.37 mmol). The reaction was stirred atroom temperature for 16 hours.

LC-MS (liquid chromatography-mass spectroscopy) showed the reaction wascompleted. The reaction mixture was concentrated in vacuum. The residuewas dissolved in water (50 ml) and acidified to pH 2-3 usingconcentrated HCl. The reaction mixture was extracted with EtOAc (2×20ml). The organic layer was washed with brine (2×50 ml), dried overanhydrous Na₂SO₄, and filtered. The filtrate was concentrated in vacuumto give the product2-((5-methoxy-2-(4-methoxy-2-pentylphenoxy)phenyl)carbamoyl)heptanoicacid (BKT300-11-a4) as a yellow solid (2.5 grams, 85.6% yield). LC-MS:m/z 472.0 (M++H).

5. Preparation of6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline-2,4(1H,3H)-dione(BKT300-11-a5)

To a PPA solution (8 ml) at 120° C. was added2-((5-methoxy-2-(4-methoxy-2-pentylphenoxy)phenyl)carbamoyl)heptanoicacid (BKT300-11-a4) (1.0 gram, 2.12 mmol) portion-wise. The reactionmixture was stirred at 120° C. for 6 hours. LC-MS (liquidchromatography-mass spectroscopy) showed the reaction was completed. Thereaction mixture was poured into water (100 ml) and extracted with EtOAc(2×20 ml). The organic layer was washed with brine (2×50 ml), dried overanhydrous Na₂SO₄, and filtered. The filtrate was concentrated in vacuum.The residue was purified by silica gel chromatography eluted with(EtOAc:Petroleum Ether=1:101:5) to give the product6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline-2,4(1H,3H)-dione(BKT300-11-a5) as a yellow solid (260 mg, 27.1% yield). LC-MS: m/z 454.7(M++H).

1H NMR (400 MHz, DMSO-d6): δ 11.76 (s, 1H), 7.56 (s, 1H), 4.05 (t, J=6.8Hz, 2H), 1.63-1.56 (m, 2H), 1.36-1.27 (m, 4H), 0.88 (t, J=6.8 Hz, 3H).

6. Preparation of2,4-dichloro-6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-11-a6)

A mixture of 6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinolone-2,4(1H,3H)-dione (BKT300-11-a5) (260 mg, 0.573 mmol) in POCl₃(2 ml) was stirred at 80° C. for 4 hours. LC-MS (liquidchromatography-mass spectroscopy) showed the reaction was completed. Thereaction mixture was concentrated in vacuum. The residue was dissolvedin EtOAc (20 ml) and washed successively with saturated NaHCO₃(2×20 ml)and brine (20 ml), dried over anhydrous Na₂SO₄, and filtered. Thefiltrate was concentrated in vacuum to give the product2,4-dichloro-6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-11-a6) as a light yellow solid (266 mg, 94.7% yield). LC-MS: m/z490.0 (M++H).

7. Preparation of4-chloro-2,6-dimethoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-11-a7)

A mixture of2,4-dichloro-6-methoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-11-a6) (71 mg, 0.144 mmol) and MeONa (76.8 mg, 14.4 mmol) intoluene (4 ml) was stirred at 145° C. in a sealed tube for 20 hours. TLCshowed the reaction was completed (EtOAc:Petroleum Ether=1:50). Thereaction mixture was concentrated in vacuum. Water was poured into theresidue and extracted with EtOAc (2×20 ml). The organic layer was washedwith brine (2×20 ml), dried over anhydrous Na₂SO₄, and filtered. Thefiltrate was concentrated in vacuum. The residue was purified by TLCeluted with (EtOAc:Petroleum Ether=1:100) to give the product4-chloro-2,6-dimethoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentyl quinoline(BKT300-11-a7) as a white solid (40 mg, 56.3% yield). LC-MS: m/z 486.0(M++H).

8. Preparation of2,6-dimethoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinoline(BKT300-11-a8)

A mixture of4-chloro-2,6-dimethoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentyl quinoline(BKT300-11-a7) (40 mg, 0.082 mmol) and 10% Pd/C (20 mg) in MeOH (20 ml)and Et₃N (2 ml) was stirred at room temperature for 2.0 hours. TLCshowed the reaction was completed (EtOAc:Petroleum Ether=1:20). Thereaction mixture was filtered. The filtrate was concentrated in vacuum.The residue was dissolved in EtOAc (20 ml) and washed with brine (2×20ml). The organic layer was dried over anhydrous Na₂SO₄, and filtered.The filtrate was concentrated in vacuum to give the product2,6-dimethoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentyl quinoline(BKT300-11-a8) as a light yellow solid (30 mg, 80.6% yield). LC-MS: m/z452.1 (M++H).

9. Preparation of6-hydroxy-8-(4-hydroxy-2-pentylphenoxy)-3-pentylquinolin-2(JH)-one(BKT300-11)

A mixture of 2,6-dimethoxy-8-(4-methoxy-2-pentylphenoxy)-3-pentylquinolone (BKT300-11-a8) (130 mg, 0.066 mmol) in 30% HBr in CH₃COOH (4ml) was stirred at 120° C. for 16 hours. Then the reaction mixture wasconcentrated in vacuum. The residue was dissolved in 30% HBr in CH₃COOH(4 ml) in a sealed tube and was stirred at 120° C. for 4 hours. Thereaction mixture was concentrated in vacuum. The residue was purified bypre-HPLC to give 6-hydroxy-8-(4-hydroxy-2-pentylphenoxy)-3-pentylquinolin-2(1H)-one (BKT300-11) as a yellow solid (36mg, yield 31.5%). LC-MS: m/z 410.8 (M++H).

1H NMR (400 MHz, CDCl₃): δ 7.99 (s, 1H), 6.42 (s, 1H), 6.40 (s, 1H),6.23 (s, 1H), 4.23 (t, J=7.6 Hz, 2H), 3.84 (s, 3H), 3.73 (s, 3H), 2.46(t, J=7.6 Hz, 2H), 1.74-1.71 (m, 2H), 1.65-1.63 (m, 1H), 1.51-1.47 (m,1H), 1.39 (m, 4H), 1.25 (m, 4H), 0.93 (t, J=6.4 Hz, 3H), 0.82 (t, J=6.4Hz, 3H).

Activity Assays:

Each of the compounds presented in Table 5 are tested in a cellmigration assay as described hereinabove, under the “methods” section,so as to determine its effect on a biological activity of the testedchemokines, and hence its activity in treating diseases associated withthe chemokine.

Of these tested compounds, compounds exhibiting modulation of abiological activity of chemokines are tested in vitro and in vivo incancer models, as described hereinabove.

Although the invention has been described in conjunction with specificembodiments thereof, it is evident that many alternatives, modificationsand variations will be apparent to those skilled in the art.Accordingly, it is intended to embrace all such alternatives,modifications and variations that fall within the spirit and broad scopeof the appended claims.

All publications, patents and patent applications mentioned in thisspecification are herein incorporated in their entirety by referenceinto the specification, to the same extent as if each individualpublication, patent or patent application was specifically andindividually indicated to be incorporated herein by reference. Inaddition, citation or identification of any reference in thisapplication shall not be construed as an admission that such referenceis available as prior art to the present invention. To the extent thatsection headings are used, they should not be construed as necessarilylimiting. In addition, any priority document(s) of this applicationis/are hereby incorporated herein by reference in its/their entirety.

What is claimed is:
 1. A method of treating an autoimmune disease in asubject in need thereof, the method comprising administering to thesubject a compound represented by the Formula:

thereby treating the autoimmune disease.
 2. The method of claim 1,wherein said autoimmune disease is a cutaneous autoimmune disease. 3.The method of claim 3, wherein said autoimmune cutaneous disease isselected from pemphigus vulgaris, bullous pemphigoid and pemphigusfoliaceus.
 4. The method of claim 1, wherein said autoimmune disease ispsoriasis.
 5. The method of claim 1, wherein the autoimmune disease isselected from an autoimmune cardiovascular disease, an autoimmunerheumatoid disease, an autoimmune glandular disease, an autoimmunegastrointestinal disease, an autoimmune hepatic disease, an autoimmuneneurological disease, an autoimmune muscular disease, an autoimmunenephric disease, an autoimmune disease related to reproduction, anautoimmune connective tissue disease and an autoimmune systemic disease.6. The method of claim 5, wherein said autoimmune cardiovascular diseaseis selected from atherosclerosis, myocardial infarction, thrombosis,Wegener's granulomatosis, Takayasu's arteritis, Kawasaki syndrome,anti-factor VIII autoimmune disease, necrotizing small vesselvasculitis, microscopic polyangiitis, Churg and Strauss syndrome,pauci-immune focal necrotizing and crescentic glomerulonephritis,antiphospholipid syndrome, antibody-induced heart failure,thrombocytopenic purpura, autoimmune hemolytic anemia, cardiacautoimmunity in Chagas' disease and anti-helper T lymphocyteautoimmunity.
 7. The method of claim 5, wherein said autoimmunerheumatoid disease is selected from rheumatoid arthritis and ankylosingspondylitis.
 8. The method of claim 5, wherein said autoimmune glandulardisease is selected from an autoimmune pancreatic disease, Type Idiabetes, an autoimmune thyroid disease, Graves' disease, thyroiditis,spontaneous autoimmune thyroiditis, Hashimoto's thyroiditis, idiopathicmyxedema, ovarian autoimmunity, autoimmune anti-sperm infertility,autoimmune prostatitis and Type I autoimmune polyglandular syndrome. 9.The method of claim 5, wherein said autoimmune gastrointestinal diseaseis selected from a chronic inflammatory intestinal disease, celiacdisease, colitis, ileitis and Crohn's disease.
 10. The method of claim5, wherein said autoimmune hepatic disease is selected from autoimmunehepatitis, autoimmune chronic active hepatitis, and primary biliarycirrhosis.
 11. The method of claim 5, wherein said autoimmuneneurological disease is selected from multiple, Alzheimer's disease,myasthenia gravis, an autoimmune neuropathy, myasthenia, Lambert-Eatonmyasthenic syndrome, a paraneoplastic neurological disease, cerebellaratrophy, paraneoplastic cerebellar atrophy, stiff-man syndrome, anon-paraneoplastic stiff man syndrome, a progressive cerebellar atrophy,encephalitis, Rasmussen's encephalitis, amyotrophic lateral sclerosis,Sydeham chorea, Gilles de la Tourette syndrome, an autoimmunepolyendocrinopathy, a dysimmune neuropathy, an acquired neuromyotonia,arthrogryposis multiplex congenita, neuritis, and optic neuritis. 12.The method of claim 5, wherein said autoimmune neurological disease is aneurodegenerative disease.
 13. The method of claim 5, wherein saidautoimmune muscular disease is selected from autoimmune myositis,primary Sjogren's syndrome and a smooth muscle autoimmune disease. 14.The method of claim 5, wherein said autoimmune nephric disease isselected from nephritis and autoimmune interstitial nephritis.
 15. Themethod of claim 5, wherein said autoimmune disease related toreproduction is repeated fetal loss.
 16. The method of claim 5, whereinsaid autoimmune connective tissue disease is selected from an autoimmuneear disease and an autoimmune disease of the inner ear.
 17. The methodof claim 5, wherein said autoimmune systemic disease is selected fromsystemic lupus erythematosus and systemic sclerosis.
 18. The method ofclaim 1, wherein said autoimmune disease is selected from Crohn'sdisease, psoriasis, scleroderma or rheumatoid arthritis.